Cephem compounds

ABSTRACT

Cephem compounds having the following formula are disclosed: ##STR1## R 1  =amino or protected amino R 2  =an organic group ##STR2## R 6  =lower alkyl R 7 , R 8  ═OH or protected OH 
     Y═N or CH 
     The compounds are useful as antimicrobial agents.

This application is a continuation of application Ser. No. 07/354,228,filed on May 19. 1989, now abandoned.

The present invention relates to new cephem compounds andpharmaceutically acceptable salts thereof. More particularly, it relatesto new cephem compounds and pharmaceutically acceptable salts thereof,which have antimicrobial activities, to processes for preparationthereof, to pharmaceutical composition comprising the same, and to amethod for treating infectious diseases in human being and animals.

Accordingly, one object of the present invention is to provide thecephem compounds and pharmaceutically acceptable salts thereof, whichare highly active against a number of pathogenic microorganisms.

Another object of the present invention is to provide processes for thepreparation of the cephem compounds and salts thereof.

A further object of the present invention is to provide pharmaceuticalcomposition comprising, as an active ingredient, said cephem compoundsor their pharmaceutically acceptable salts.

Still further object of the present invention is to provide a method fortreating infectious diseases caused by pathogenic microorganisms, whichcomprises administering said cephem compounds to infected human being oranimals.

The object cephem compounds are novel and can be represented by thefollowing general formula (I): ##STR3## wherein R¹ is amino or aprotected amino group, Z is N or CH,

R² is an organic group, and

R is a group of the formula: ##STR4##

(in which R³ and R⁴ are each lower alkyl, or

R³ and R⁴ are linked together to form C₃ -C₆ alkylene,

A is lower alkylene, and

R⁵ is hydroxy or a protected hydroxy group)

or a group of the formula: ##STR5## (in which R⁶ is lower alkyl, R⁷ andR⁸ are each hydroxy or a protected hydroxy group, and

Y is N or CH).

The cephem compound (I) of the present invention can be prepared byprocesses as illustrated in the following. ##STR6## wherein R, R¹, R²,R⁶, R⁷, R⁸, Z and Y are each as defined above,

X is an acid residue,

R^(a) is a compound of the formula: ##STR7## (in which R³, R⁴, R⁵ and Aare each as defined above) or a compound of the formula: ##STR8## (inwhich R⁶, R⁷, R⁸ and Y are each as defined above), R_(a) ¹ is aprotected amino group,

R_(a) ² is protected carboxy(lower)alkyl,

R_(b) ² is carboxy(lower)alkyl, and

R_(a) ⁸ is a protected hydroxy group.

Some of the starting compounds are novel and can be prepared byprocesses as illustrated in the following. ##STR9## wherein R¹, R², R³,R⁴, R⁵, R⁶, R⁷, R⁸, R_(a) ⁸ A, X, Y, Z, R and R^(a) are each as definedabove,

W acid residue,

R_(a) ⁵ is a protected hydroxy group,

R⁹ is amino or a protected amino group,

R_(a) ⁹ is a protected amino group, and

R¹⁰ is an imino protective group.

Regarding the compounds (I), (Ia)˜(Ig), (II), (V), (VI) and (XIV), it isto be understood that said compounds include syn isomer, anti isomer anda mixture thereof.

For example, with regard to the object compounds (I), syn isomer meansone geometrical isomer having the partial structure represented by thefollowing formula: ##STR10## (wherein R¹, R² and Z are each as definedabove) and anti isomer means the other geometrical isomer having thepartial structure represented by the following formula: ##STR11##(wherein R¹, R² and Z are each as defined above), and all of suchgeometrical isomers and mixture thereof are included within the scope ofthis invention.

In the present specification and claim, the partial structure of thesegeometrical isomers and mixture thereof are represented for convenientsake by the following formula: ##STR12## (wherein R¹, R² and Z are eachas defined above).

Further, regarding the compounds (I), (Ia)˜(Ig), (III), (IIIe), (IIIf),(IIIg), (IV), (IVa), (IVb) and (VII), it is to be understood that thesaid compounds include tautomeric isomers. For example, with regard tothe object compound (I), in case that the symbol "R" in the compound (I)means the group represented by the following formula: ##STR13## (whereinR⁶ and R⁸ are each as defined above), said group can also exist in thetautomeric form and such tautomeric equilibrium can be represented bythe following scheme. ##STR14##

Both of the above tautomeric isomers are included within the scope ofthe present invention. In the present specification and claim, thecompounds including the group of such tautomeric isomers are representedfor the convenient sake by one expression of the group of the formula(A).

Still further, regarding the compounds (I), (Ia)˜(Id), (III), (IIIa),(IIIb), (IIIc), (IIId), (IV), (IVa), (IVb), (IX) and (X), it is to beunderstood that the said compounds include tautomeric isomers. Forexample, with regard to the object compound (I), in case that the symbol"R" in the compound (I) means the group represented by the followingformula: ##STR15## (wherein R³, R⁴, R⁵ and A are each as defined above),said group can also exist in the tautomeric form and such tautomericequilibrium and be represented by the following scheme. ##STR16##

Both the of the above tautomeric isomers are included within the scopeof the present invention. In the present specification and claim, thecompounds including the group of such tautomeric isomers are representedfor the convenient sake by one expression of the group of the formula(A').

In the above and subsequent descriptions of the present specification,suitable examples and illustration of the various definitions which thepresent invention include within the scope thereof are explained indetail as follows.

The term "lower" is intended to mean 1 to 6 carbon atom(s) unlessotherwise indicated.

Suitable "protected amino" may include an acylamino or an amino groupsubstituted by a conventional protecting group such as ar(lower)alkylwhich may have suitable substituent(s) (e.g. benzyl, trityl, etc.) orthe like.

Suitable "acyl moiety" in the term "acylamino" may include carbamoyl,aliphatic acyl group and acyl group containing an aromatic orheterocyclic ring. And, suitable examples of the said acyl may be loweralkanoyl (e.g. formyl, acetyl, propionyl, butyryl, isobutyryl, valeryl,isovaleryl, oxalyl, succinyl, pivaloyl, etc.); lower alkoxycarbonyl(e.g. methoxycarbonyl, ethoxycarbonyl, propoxycarbonyl,1-cyclopropylethoxycarbonyl, isopropoxycarbonyl, butoxycarbonyl,tertiarybutoxycarbonyl, pentyloxycarbonyl, hexyloxycarbonyl, etc.);lower alkanesulfonyl (e.g. mesyl, ethanesulfonyl, propanesulfonyl,isopropanesulfonyl, butanesulfonyl, etc.); arenesulfonyl (e.g.benzenesulfonyl, tosyl, etc.); aroyl (e.g. benzoyl, toluoyl, xyloyl,naphthoyl, phthaloyl, indancarbonyl, etc.); ar(lower)alkanoyl (e.g.phenylacetyl, phenylpropionyl, etc.); ar(lower)alkoxycarbonyl (e.g.benzyloxycarbonyl, phenethyloxycarbonyl, etc.), and the like. The acylmoiety as stated above may have suitable substituent(s) such as halogen(e.g. chlorine, bromine, iodine or fluorine) or the like.

Suitable "organic group" may include lower alkyl (e.g., methyl, ethyl,propyl, isopropyl, butyl, isobutyl, tert-butyl, pentyl, neopentyl,tert-pentyl, hexyl, etc.),

mono(or di or tri)halo[lower)alkyl (e.g. chloromethyl, dichloromethyl,trichloromethyl, bromomethyl, chloroethyl, dichloroethyl,trichloroethyl, fluoroethyl, trifluoroethyl, etc.),

lower alkenyl (e.g., vinyl, 1-propenyl, allyl, 1-methylallyl, 1 or 2 or3-butenyl, 1 or 2 or 3 or 4-pentenyl, 1 or 2 or 3 or 4 or 5-hexenyl,etc.),

lower alkynyl (e.g., ethynyl, 1-propynyl, propargyl, 1-methylpropargyl,1 or 2 or 3 butynyl, 1 or 2 or 3 or 4-pentynyl, 1 or 2 or 3 or 4 or5-hexynyl, etc.),

aryl (e.g., phenyl, naphthyl, etc.),

ar(lower)alkyl such as phenyl(lower)alkyl (e.g., benzyl, phenethyl,phenylpropyl, etc.),

carboxy(lower)alkyl wherein lower alkyl moiety can be referred to theones as exemplified above, protected carboxy(lower)alkyl wherein loweralkyl moiety can be referred to the ones as exemplified above andprotected carboxy moiety can be referred to the ones as exemplifiedbelow, and the like.

Suitable "protected carboxy moiety" in the term "protectedcarboxy(lower)alkyl" may include esterified carboxy and the like. Andsuitable examples of said ester may be the ones such as lower alkylester (e.g., methyl ester, ethyl ester, propyl ester, isopropyl ester,butyl ester, isobutyl ester, t-butyl ester, pentyl ester, t-pentylester, hexyl ester, 1-cyclopropylethyl ester, etc.); lower alkenyl ester(e.g., vinyl ester, allyl ester, etc.); lower alkynyl ester (e.g.,ethynyl ester, propynyl ester, etc.); lower alkoxyalkyl ester (e.g.,methoxymethyl ester, ethoxymethyl ester, isopropoxymethyl ester,1-methoxyethyl ester, 1-ethoxyethyl ester, etc.); lower alkylthioalkylester (e.g., methylthiomethyl ester, ethylthiomethyl ester,ethylthioethyl ester, isopropylthiomethyl ester, etc.); mono(or di ortri)halo(lower)alkyl ester (e.g. 2-iodoethyl ester, 2,2,2-trichloroethylester, etc.); lower alkanoyloxy(lower)alkyl ester (e.g., acetoxymethylester, propionyloxymethyl ester, butyryloxymethyl ester,valeryloxymethyl ester, pivaloyloxymethyl ester, hexanoyloxymethylester, 2-acetoxyethyl ester, 2-propionyloxyethyl ester, etc.); loweralkanesulfonyl(lower)alkyl ester (e.g. mesylmethyl ester, 2-mesylethylester etc.); ar(lower)alkyl ester, for example, phenyl(lower)alkyl esterwhich may have one or more suitable substituent(s) (e.g., benzyl ester,4-methoxybenzyl ester, 4-nitrobenzyl ester, phenethyl ester, tritylester, benzhydryl ester, bis(methoxyphenyl)methyl ester,3,4-dimethoxybenzyl ester, 4-hydroxy-3,5-di-t-butylbenzyl ester, etc.);aryl ester which may have one or more suitable substituent(s) such assubstituted or unsubstituted phenyl ester (e.g., phenyl ester, tolylester, t-butylphenyl ester, xylyl ester, mesityl ester, cumenyl ester,4-chlorophenyl ester, 4-methoxyphenyl ester, etc.); tri(lower)alkylsilyl ester; lower alkylthioester (e.g. methylthioester, ethylthioester,etc.) and the like.

Suitable "lower alkyl" can be referred to the ones as exemplified above.

Suitable "C₃ -C₆ alkylene" may include trimethylene, tetramethylene,pentamethylene and hexamethylene.

Suitable "lower alkylene" may include methylene, ethylene, trimethylene,tetramethylene, pentamethylene, hexamethylene and the like.

Suitable "protective group" in the "protected hydroxy group" may includeacyl as mentioned above, tetrahydropyranyl and the like.

Suitable "acid residue" may include halogen [e.g. chlorine, bromine,iodine, etc.], acyloxy such as sulfonyloxy [e.g. benzenesulfonyloxy,tosyloxy, mesyloxy, etc.], lower alkanoyloxy [e.g. acetyloxy,propionyloxy, etc.] or the like.

Suitable "imino protective group" may include lower alkoxycarbonyl[e.g., methoxycarbonyl, ethoxycarbonyl, propoxycarbonyl,isopropoxycarbonyl, butoxycarbonyl, tert-butoxycarbonyl,pentyloxycarbonyl, tert-pentyloxycarbonyl, hexyloxycarbonyl, etc.]andthe like.

Suitable pharmaceutically acceptable salts of the object compound (I)are conventional non-toxic salts and include a metal salt such as analkali metal salt [e.g. sodium salt, potassium salt, etc.] and analkaline earth metal salt [e.g. calcium salt, magnesium salt, etc.], anammonium salt, an organic base salt [e.g. trimethylamine salt,triethylamine salt, pyridine salt, picoline salt, dicyclohexylaminesalt, N,N'-dibenzylethylenediamine salt, etc], an organic acid salt[e.g. formate, acetate, trifluoroacetate, maleate, tartrate,methanesulfonate, benzenesulfonate, toluenesulfonate, etc.], aninorganic acid salt [e.g. hydrochloride, hydrobromide, sulfate,phosphate, etc.], a salt with an amino acid [e.g. arginine salt,aspartic acid salt, glutamic acid salt, etc.], and the like.

Preferred embodiments of the object compound (I) are as follows.

R¹ is amino or acylamino (more preferably lower alkanoylamino),

Z is N or CH,

R² is lower alkyl, lower alkenyl, carboxy(lower)alkyl or protectedcarboxy(lower)alkyl [more preferably esterified carboxy(lower)alkyl,most preferably lower alkoxycarbonyl(lower)alkyl],

R³ and R⁴ are each lower alkyl, or

R³ and R⁴ are linked together to form C₃ -C₆ alkylene,

A is lower alkylene (more preferably C₁ -C₃ alkylene),

R⁵ is hydroxy,

R⁶ is lower alkyl,

R⁷ is hydroxy or acyloxy (more preferably lower alkanoyloxy),

R⁸ is hydroxy or acyloxy (more preferably lower alkanoyloxy), and

Y is N or CH.

The processes for preparing the object and starting compounds of thepresent invention are explained in detail in the following.

Process 1

The compound (I or a salt thereof can be prepared by reacting thecompound (II) or a salt thereof with the compound (III) or a saltthereof.

Suitable salts of the compounds (II) and (III) can be referred to theones as exemplified for the compound (I).

The present reaction may be carried out in a solvent such as water,phosphate buffer, acetone, chloroform, acetonitrile, methylene chloride,ethylene chloride, formamide, N,N-dimethylformamide, methanol, ethanol,diethyl ether, tetrahydrofuran, dimethyl sulfoxide, or any other organicsolvent which does not adversely affect the reaction. Among thesolvents, hydrophilic solvents may be used in a mixture with water. Thereaction temperature is not critical, and the reaction is usuallycarried out under cooling, at ambient temperature or under warming.

Process 2

The compound (Ib) or a salt thereof can be prepared by subjecting thecompound (Ia) or a salt thereof to elimination reaction of the aminoprotective group.

Suitable salts of the compounds (Ia) and (Ib) can be referred to theones as exemplified for the compound (I).

This reaction is carried out in accordance with a conventional methodsuch as hydrolysis, reduction or the like.

The hydrolysis is preferably carried out in the presence of a base or anacid including Lewis acid. Suitable base may include an inorganic baseand an organic base such as an alkali metal [e.g. sodium, potassium,etc.], an alkaline earth metal [e.g. magnesium, calcium, etc.], thehydroxide or carbonate or bicarbonate thereof, trialkylamine [e.g.trimethylamine, triethylamine, etc.], picoline,1,5-diazabicyclo[4.3.0]non-5-ene, 1,4-diazabicyclo[2.2.2]octane,1,8-diazabicyclo[5.4.0]-undec-7-ene, or the like.

Suitable acid may include an organic acid [e.g. formic acid, aceticacid, propionic acid, trichloroacetic acid, trifluoroacetic acid, etc.]and an inorganic acid [e.g. hydrochloric acid, hydrobromic acid,sulfuric acid, hydrogen chloride, hydrogen bromide, etc.]. Theelimination using Lewis acid such as trihaloacetic acid [e.g.trichloroacetic acid, trifluoroacetic acid, etc.] or the like ispreferably carried out in the presence of cation trapping agents [e.g.anisole, phenol, etc.].

The reaction is usually carried out in a solvent such as water, analcohol [e.g. methanol, ethanol, etc.], methylene chloride,tetrahydrofuran, a mixture thereof or any other solvent which does notadversely affect the reaction. A liquid base or acid can be also used asthe solvent. The reaction temperature is not critical and the reactionis usually carried out under cooling to warming.

The reduction method applicable for the elimination reaction may includechemical reduction and catalytic reduction.

Suitable reducing agents to be used in chemical reduction are acombination of metal [e.g. tin, zinc, iron, etc.] or metallic compound[e.g. chromium chloride, chromium acetate, etc.]and an organic orinorganic acid [e.g. formic acid, acetic acid, propionic acid,trifluoroacetic acid, p-toluenesulfonic acid, hydrochloric acid,hydrobromic acid, etc.].

Suitable catalysts to be used in catalytic reduction are conventionalones such as platinum catalysts [e.g. platinum plate, spongy platinum,platinum black, colloidal platinum, platinum oxide, platinum wire,etc.], palladium catalysts [e.g. spongy palladium, palladium black,palladium oxide, palladium on carbon, colloidal palladium, palladium onbarium sulfate, palladium on barium carbonate, etc.], nickel catalysts[e.g. reduced nickel, nickel oxide, Raney nickel, etc.], cobaltcatalysts [e.g. reduced cobalt, Raney cobalt, etc.], iron catalysts[e.g. reduced iron, Raney iron, etc.], copper catalysts [e.g. reducedcopper, Raney copper, Ullman copper, etc.] and the like.

The reduction is usually carried out in a conventional solvent whichdoes not adversely affect the reaction such as water, methanol, ethanol,propanol, N,N-dimethylformamide, or a mixture thereof. Additionally, incase that the abovementioned acids to be used in chemical reduction arein liquid, they can also be used as a solvent, further, a suitablesolvent to be used in catalytic reduction may be the above-mentionedsolvent, and other conventional solvent such as diethyl ether, dioxane,tetrahydrofuran, alcohol (e.g. methanol, ethanol, etc.), etc., or amixture thereof.

The reaction temperature of this reduction is not critical and thereaction is usually carried out under cooling to warming.

Process 3

The compound (Id) or a salt thereof can be prepared by subjecting thecompound (Ic) or a salt thereof to elimination reaction of the carboxyprotective group.

Suitable salts of the compound (Ic) and (Id) can be referred to the onesas exemplified for the compound (I).

This elimination can be carried out in a similar manner to that of theaforementioned Process 2, and therefore the reagents to be used and thereaction conditions (e.g. solvent, reaction temperature, etc.) can bereferred to those of the Process 2.

Process 4

The compound (I) or a salt thereof can be prepared by reacting thecompound (IVb) or its reactive derivative at the amino group or a saltthereof with the compound (V) or its reactive derivative at the carboxygroup or a salt thereof.

Suitable reactive derivative at the amino group of the compound (IVb)may include Schiff's base type imino or its tautomeric enamine typeisomer formed by the reaction of the compound (IVb) with a carbonylcompound such as aldehyde, ketone or the like; a silyl derivative formedby the reaction of the compound (IVb) with a silyl compound such asbis(trimethylsilyl)acetamide, mono(trimethylsilyl)acetamide [e.g.N-(trimethylsilyl)acetamide], bis(trimethylsilyl)urea or the like; aderivative formed by reaction of the compound (IVb) with phosphorustrichloride or phosgene, and the like.

Suitable salts of the compound (IVb) and its reactive derivative can bereferred to the ones as exemplified for the compound (I).

Suitable reactive derivative at the carboxy group of the compound (V)may include an acid halide, an acid anhydride, an activated amide, anactivated ester, and the like. Suitable examples of the reactivederivatives may be an acid chloride; an acid azide; a mixed acidanhydride with an acid such as substituted phosphoric acid [e.g.dialkylphosphoric acid, phenylphosphoric acid, diphenylphosphoric acid,dibenzylphosphoric acid, halogenated phosphoric acid, etc.],dialkylphosphorous acid, sulfurous acid, thiosulfuric acid, sulfuricacid, sulfonic acid [e.g. methanesulfonic acid, etc.], aliphaticcarboxylic acid e.g. acetic acid, propionic acid, butyric acid,isobutyric acid, pivalic acid, pentanoic acid, isopentanoic acid,2-ethylbutyric acid, trichloroacetic acid, etc.]or aromatic carboxylicacid [e.g. benzoic acid, etc.]; a symmetrical acid anhydride; anactivated amide with imidazole, 4-substituted imidazole,dimethylpyrazole, triazole or tetrazole; or an activated ester [e.g.cyanomethyl ester, methoxymethyl ester, dimethyliminomethyl [(CH₃)₂ N³⁰═CH--] ester, vinyl ester, propargyl ester, p-nitrophenyl ester,2,4-dinitrophenyl ester, trichlorophenyl ester, pentachlorophenyl ester,mesylphenyl ester, phenylazophenyl ester, phenyl thioester,p-nitrophenyl thioester, p-cresyl thioester, carboxymethyl thioester,pyranyl ester, pyridyl ester, piperidyl ester, 8-quinolyl thioester,etc.], or an ester with a N-hydroxy compound [e.g.N,N-dimethylhydroxylamine, 1-hydroxy-2-(1H)-pyridone,N-hydroxysuccinimide, N-hydroxyphthalimide, 1-hydroxy-1H-benzotriazole,etc.], and the like. These reactive derivatives can optionally beselected from them according to the kind of the compound (V) to be used.

Suitable salts of the compound (V) and its reactive derivative can bereferred to the ones as exemplified for the compound (I).

The reaction is usually carried out in a conventional solvent such aswater, alcohol [e.g. methanol, ethanol, etc.], acetone, dioxane,acetonitrile, chloroform, methylene chloride, ethylene chloride,tetrahydrofuran, ethyl acetate, N,N-dimethylformamide, pyridine or anyother organic solvent which does not adversely influence the reaction.These conventional solvent may also be used in a mixture with water.

In this reaction, when the compound (V) is used in a free acid form orits salt form, the reaction is preferably carried out in the presence ofa conventional condensing agent such as N,N'-dicyclohexylcarbodiimide;N-cyclohexyl-N'-morpholinoethylcarbodiimide;N-cyclohexyl-N'-(4-diethylaminocyclohexyl)carbodiimide;N,N'-diethylcarbodiimide, N,N'-diisopropylcarbodiimide;N-ethyl-N'-(3-dimethylaminopropyl)carbodiimide;N,N'-carbonylbis-(2-methylimidazole);pentamethyleneketene-N-cyclohexylimine;diphenylketene-N-cyclohexylimine; ethoxyacetylene;1-alkoxy-1-chloroethylene; trialkyl phosphite; ethyl polyphosphate;isopropyl polyphosphate; phosphorus oxychloride (phosphoryl chloride);phosphorus trichloride; thionyl chloride; oxalyl chloride; lower alkylhaloformate [e.g. ethyl chloroformate, isopropyl chloroformate, etc.];triphenylphosphine; 2-ethyl-7-hydroxybenzisoxazolium salt;2-ethyl-5-(m-sulfophenyl)isoxazolium hydroxide intramolecular salt;1-(p-chlorobenzenesulfonyloxy)-6-chloro-1H-benzotriazole; so-calledVilsmeier reagent prepared by the reaction of N,N-dimethylformamide withthionyl chloride, phosgene, trichloromethyl chloroformate, phosphorusoxychloride, etc.; or the like.

The reaction may also be carried out in the presence of an inorganic ororganic base such as an alkali metal bicarbonate, tri(lower)alkylamine,pyridine, N-(lower alkylmorpholine, N,N-di(lower)alkylbenzylamine, orthe like.

The reaction temperature is not critical, and the reaction is usuallycarried out under cooling to warming.

Process 5

The compound (Ie) or a salt thereof can be prepared by reacting thecompound (VI) or its reactive derivative at the imino group or a saltthereof with the compound (VII) or its reactive derivative at thecarboxy group or a salt thereof.

Suitable reactive derivative at the imino group of the compound (VI) mayinclude a silyl derivative formed by the reaction of the compound (VI)with a silyl compound (e.g., N,O-bis(trimethylsilyl)acetamide,N-trimethylsilylacetamide, etc. ; and the like.

Suitable salts of the compound (VI) and (VII) can be referred to theones as exemplified for the compound (I).

Suitable reactive derivative at the carboxy group of the compound (VII)may include an acid halide, an acid anhydride, an activated amide, anactivated ester, and the like.

This reaction can be carried out in a similar manner to that of theaforementioned Process 4, and therefore the reagents to be used and thereaction conditions (e.g. solvent, reaction temperature, etc.) can bereferred to those of the Process 4.

Process 6

The compound (Ig) or a salt thereof can be prepared by subjecting thecompound (If) or a salt thereof to elimination reaction of the hydroxyprotective group.

Suitable salts of the compounds (If) and (Ig) can be referred to theones as exemplified for the compound (I).

This elimination can be carried out in a similar manner to that of theaforementioned Process 2, and therefore the reagents to be used and thereaction conditions (e.g. solvent, reaction temperature, etc.) can bereferred to those of the Process 2.

Process A

The compound (IIIa) or a salt thereof can be prepared by reacting thecompound (VIII) or a salt thereof with the compound (IX) or a saltthereof.

The present reaction is usually carried out in the presence of a base.

Suitable base can be referred to the ones as exemplified in Process 2.

The reaction is usually carried out in a solvent such as water, analcohol [e.g. methanol, ethanol, etc.], methylene chloride,tetrahydrofuran, a mixture thereof or any other solvent which does notadversely affect the reaction. The reaction temperature is not criticaland the reaction is usually carried out under cooling to warming.

Process B

The compound (IIIb) or a salt thereof can be prepared by reacting thecompound (VIII) or a salt thereof with the compound (X) or a saltthereof.

The reaction is usually carried out in the presence or absence of asolvent such as water, an alcohol [e.g. methanol, ethanol, etc.],methylene chloride, tetrahydrofuran, a mixture thereof or any othersolvent which does not adversely affect the reaction. The reactiontemperature is not critical and the reaction is usually carried outunder warming to heating.

Process C

The compound (IIId) or a salt thereof can be prepared by subjecting thecompound (IIIc) or a salt thereof to elimination reaction of the hydroxyprotective group.

The elimination reaction can be carried out in a similar manner to thatof the aforementioned Process 2, and therefore the reagents to be usedand the reaction conditions (e.g. solvent, reaction temperature, etc.)can be referred to those of the Process 2.

Process D

The compound (IV) or a salt thereof can be prepared by reacting thecompound (XI) or a salt thereof with the compound (III) or a saltthereof.

This reaction can be carried out in a similar manner to that of theaforementioned Process 1, and therefore the reagents to be used and thereaction conditions (e.g. solvent, reaction temperature, etc.) can bereferred to those of the Process 1.

Process E

The compound (IVb) or a salt thereof can be prepared by subjecting thecompound (IVa) or a salt thereof to elimination reaction of the aminoprotective group.

This elimination can be carried out in a similar manner to that of theaforementioned Process 2, and therefore the reagents to be used and thereaction conditions (e.g. solvent, reaction temperature, etc.) can bereferred to those of the Process 2.

Process F

The compound (IIIe) or a salt thereof can be prepared by reacting thecompound (XII) or its reactive derivative at the imino group or a saltthereof with the compound (VII) or its reactive derivative at thecarboxy group or a salt thereof.

Suitable reactive derivative at the imino group of the compound (XII)may include the same one as exemplified for the compound (VI) in Process5.

The present reaction can be carried out in a similar manner to that ofthe aforementioned Process 4, and therefore the reagents to be used andthe reaction conditions (e.g. solvent, reaction temperature, etc.) canbe referred to those of the Process 4.

Process G

The compound (IIIg) or a salt thereof can be prepared by subjecting thecompound (IIIf) or a salt thereof to elimination reaction of the hydroxyprotective group.

This elimination reaction can be carried out in a similar manner to thatof the aforementioned Process 2, and therefore the reagents to be usedand the reaction conditions (e.g. solvent, reaction temperature, etc.)can be referred to those of the Process 2.

Process H--1

The compound (XIV) or a salt thereof can be prepared by reacting thecompound (II) or a salt thereof with the compound (XIII).

The present reaction can be carried out in a similar manner to that ofthe aforementioned Process 1, and therefore the reagent to be used andthe reaction conditions (e.g. solvent, reaction temperature, etc.) canbe referred to those of the Process 1.

Process H--2

The compound (VI) or a salt thereof can be prepared by subjecting thecompound (XIV) or a salt thereof to elimination reaction of the iminoprotective group.

This elimination can be carried out in a similar manner to that of theaforementioned Process 2, and therefore the reagents to be used and thereaction conditions (e.g. solvent, reaction temperature, etc.) can bereferred to those of the Process 2.

The object compound (I) and pharmaceutically acceptable salts thereofare novel and exhibit high antimicrobial activity, inhibiting the growthof a wide variety of pathogenic microorganisms including Gram-positiveand Gram-negative microorganisms and are useful as antimicrobial agents.

Now in order to show the utility of the object compound (I), the testdata on MIC (minimal inhibitory concentration) of representativecompound of this invention are shown in the following.

Test method

In vitro antibacterial activity was determined by the two-foldagar-plate dilution method as described below.

One loopful of an overnight culture of each test strain inTrypticase-soy broth (10⁸ viable cells per ml) was streaked on heartinfusion agar (HI-agar)containing graded concentrations ofrepresentative test compound, and the minimal inhibitory concentration(MIC) was expressed in terms of λg/ml after incubation at 37° C. for 20hours.

Test compound

(1)7β-[2-(5-Amino-1,2,4-thiadiazol-3-yl)-2-(1-carboxy-1-methylethoxyimino)acetamido]-3-[N,N-dimethyl-N-{(5-hydroxy-4-oxo-1,4-dihydropyridin-2-yl)methyl}-ammonio]methyl-3-cephem-4-carboxylate(syn isomer)

(2)7β-[2-(5-Amino-1,2,4-thiadiazol-3-yl)-2-methoxyiminoacetamido]-3-[1-methyl-4-(4,5-dihydroxy-2-pyridylcarbonyl)-1-piperazinio]methyl-3-cephem-4-carboxylate(syn isomer)

    ______________________________________                                        Test result                                                                   MIC (μg/ml)                                                                               Test compounds                                                 Test strain      (1)     (2)                                                  ______________________________________                                        P. aeruginosa 26 ≦0.025                                                                         0.05                                                 ______________________________________                                    

For therapeutic administration, the object compound (I) andpharmaceutically acceptable salts thereof of the present invention arused in the form of conventional pharmaceutical preparation whichcontains said compound as an active ingredient, in admixture withpharmaceutically acceptable carriers such as an organic or inorganicsolid or liquid excipient which is suitable for oral, parenteral andexternal administration.

The pharmaceutical preparations may be in solid form such as tablet,granule, powder, capsule, or liquid form such as solution, suspension,syrup, emulsion, lemonade and the like.

If needed, there may be included in the above preparations, auxiliarysubstances, stabilizing agents, wetting agents and other commonly usedadditives such as lactose, citric acid, tartaric acid, stearic acid,magnesium stearate, terra alba, sucrose, corn starch, talc, gelatin,agar, pectin, peanut oil, olive oil, cacao butter, ethylene glycol, andthe like.

While the dosage of the compound (I) may vary from and also depend uponthe age, conditions of the patient, a kind of diseases, a kind of thecompound (I) to be applied, etc., in general, amounts between 1 mg andabout 4,000 mg or even more per day may be administered to a patient. Anaverage single dose of about 50 mg, 100 mg, mg, 500 mg, 1000 mg of theobject compound (I) of the present invention may be used in treatingdiseases infected by pathogenic microorganisms.

The following Preparations and Examples are given for the purpose ofillustrating the present invention in more detail.

Preparation 1

To a mixture of 2-chloromethyl-5-benzyloxy-4-pyridone (1.0 g),tetrahydrofuran (10 ml) and water (10 ml) were added dimethylaminehydrochloride (1.31 g) and sodium hydroxide (0.64 g). After beingstirred for 1.5 hours, the mixture was concentrated under reducedpressure to dryness. The residue was dissolved in methanol and theinsoluble material was filtered off. The filtrate was evaporated invacuo to give 5-benzyloxy-2-(N,N-dimethylamino)methyl-4-pyridone (1.0 g)as a powder.

IR (Nujol) : 1620 cm⁻¹.

NMR (DMSO-d₆, δ) 2.20 (6H, s), 3.36 (2H, s), 5.06 (2H, s), 6.28 (1H, s),7.1 (5H, m), 7.13 (1H, s).

Preparation 2

A solution of 5-benzyloxy-2-chloromethyl-4-pyridone (10.0 g) andtriphenylphosphine (10.5 g) in N,N-dimethylformamide (50 ml) was stirredfor 5 hours at 90°-100° C. The resulting mixture was poured into ethylacetate (800 ml). The precipitate was collected by filtration, washedwith ethyl acetate and dissolved in dichloromethane (500 ml). To thesolution were added water (300 ml) and 38% aqueous formaldehyde (100ml). The mixture was adjusted to pH 10-10.5 with potassium carbonate.After being stirred for 3 hours at 35°-40° C., the organic layer wasseparated, washed with water, dried over magnesium sulfate andconcentrated under reduced pressure. The product was isolated by columnchromatography on silica gel with ethyl acetate as an eluent to give5-benzyloxy-2-vinyl-4-pyridone (6.02 g).

IR (Nujol):1640, 1615 cm⁻¹.

NMR (DMSO-d₆, δ) 5.10 (2H, s), 5.46 (1H, d, J=10Hz), 5.95 (1H, d,J=18Hz), 6.57 (1H, s), 6.63 (1H, dd, J=10Hz, 18Hz), 7.43 (5H, m), 7.66(1H, s).

Preparation 3

A mixture of 5-benzyloxy-2-vinyl-4-pyridone (2.90 g) and pyrrolidine(5.33 ml) was heated and refluxed for an hour. The mixture was cooledand diluted with tetrahydrofuran (20 ml) and diisopropyl ether (80 ml).After being stirred for an hour at ambient temperature, the resultingprecipitate was collected by filtration, washed with diisopropyl etherand air-dried at ambient temperature to give5-benzyloxy-2-[2-(1-pyrrolidinyl)ethyl]4-pyridone (3.79 g).

IR (Nujol) : 1633, 1618 cm⁻¹.

NMR (DMSO-d₆, δ) 1.70 (4H, m), 2.3-2.7 (8H, m), 5.03 (2H, s), 6.15 (1H,s), 7.40 (5H, m), 7.41 (1H, s).

Preparation 4

A mixture of 5-benzyloxy-2-vinyl-4-pyridone (3.12 g), 50% aqueoussolution of dimethylamine (15 ml) and ethanol (35 ml) was heated at 100°C. in a sealed tube for 8 hours. The resulting mixture was cooled andconcentrated under reduced pressure to dryness. The residue wastriturated with a mixture of ethyl acetate and diisopropyl ether to give5-benzyloxy-2-[2-(N,N-dimethylamino)ethyl]-4-pyridone (3.45 g) as apowder.

IR (Nujol):1620 (sh), 1613 cm⁻¹.

NMR (DMSO-d₆, δ) 2.16 (6H, m), 2.55 (2H, m), 5.01 (2H, s), 6.13 (1H, s),7.38 (5H, m), 7.40 (1H, s).

Preparation 5

(1) 5-Benzyloxy-2-(N,N-dimethylamino)methyl-4-pyridone 1.0 g) inmethanol (15 ml) was subjected to catalytic reduction with 10% palladiumon activated carbon (200 mg) at atmospheric pressure. After removal ofcatalyst, the solution was concentrated under reduced pressure to give2-(N,N-dimethylamino)methyl-5-hydroxy-4-pyridone (0.96 g).

IR (Nujol):3300 (br), 1620 cm⁻¹.

NMR (DMSO-d₆, δ) 2.27 (6H, s), 3.43 (2H, s), 6.30 (1H, s), 7.40 (1H, s).

The following compounds were obtained according to a similar manner tothat of Preparation 5(1).

(2) 5-Hydroxy-2-[2-(N,N-dimethylamino)ethyl]-4-pyridone

IR (Nujol):1640 (sh), 1630 cm⁻¹.

NMR (D₂ O, δ) 2.50 (6H, m), 2.92 (2H, m), 6.48 (1H, s), 7.47 (1H, s).

(3) 5-Hydroxy-2-[2-(1-pyrrolidinyl)ethyl]-4-pyridone

IR (Nujol):1623 (sh), 1608 cm⁻¹.

NMR (D₂ O, δ) 1.8-2.1 (4H, m), 2.7-3.3 (8H, m), 6.43 (1H, s), 7.36 (1H,s).

EXAMPLE 1

(1) To a mixture of benzhydryl78-[2-(5-amino-1,2,4-thiadiazol-3-yl)-2-methoxyiminoacetamido]-3-chloromethyl-3-cephem-4-carboxylate(syn isomer) (1.47 g), dichloromethane (5 ml) and anisole (1.4 ml) wasadded trifluoroacetic acid (5 ml) under ice-cooling with stirring. Afterbeing stirred for 30 minutes at the same temperature, the mixture waspoured into diisopropyl ether (100 ml). The resulting precipitate wascollected by filtration, washed with diisopropyl ether and dried underreduced pressure to give 7-[2-(5-amino-1,2,4-thiadiazol-3-yl)-2-methoxyiminoacetamido]3-chloromethyl-3-cephem-4-carboxylicacid trifluoroacetate (syn isomer). This compound was dissolved inN,N-dimethylformamide (15 ml). To the solution was added2-(N,N-dimethylamino)-methyl-5-hydroxy-4-pyridone (1.24 g). After beingstirred for 5 hours at ambient temperature, the reaction mixture waspoured into ethyl acetate (100 ml). The resulting precipitate wascollected by filtration, washed with ethyl acetate. The precipitate wassuspended in water (100 ml) and the suspension was adjusted to pH 2.0with diluted hydrochloric acid. After removal of insoluble material byfiltration, the aqueous solution was subjected to column chromatographyon Diaion HP-20 [Trademark:prepared by Mitsubishi Chemical Industries].The column was washed with water and the elution was carried out with30% aqueous methanol. The fractions containing desired product werecombined and methanol was evaporated in vacuo. The resulting aqueouslayer was lyophilized to give7β-[2-(5-amino-1,2,4-thiadiazol3-yl)-2-methoxyiminoacetamido]-3-[N,N-dimethyl-N-{(5-hydroxy-4-oxo-1,4-dihydropyridin-2-yl)methyl}-ammonio]methyl-3-cephem-4-carboxylate(syn isomer) (0.28 g) as a powder.

IR (Nujol):3250 (br), 1765, 1650, 1600 cm⁻¹.

NMR (D₂ O-NaHCO₃, δ) 3.0 (3H, br.s), 3.07 (3H, br.s), 3.40, 3.71 (2H,ABq, J=17Hz), 3.85-4.9 (4H, m), 4.05 (3H, s), 5.33 (1H, d, J=5Hz), 5.85(1H, d, J=5Hz), 6.73 (1H, s), 7.70 (1H, s) .

The following compounds were obtained according to a similar manner tothat of Example 1(1).

(2)7β-[2-(2-Formamidothiazol-4-yl)-2-tert-butoxycarbonylmethoxyiminoacetamido]-3-[N,N-dimethyl-N-{(5-hydroxy-4-oxo-1,4-dihydropyridin-2-yl)methyl}-ammonio]methyl-3-cephem-4-carboxylate(syn isomer)

IR (Nujol):3300 (br), 1780, 1730, 1675, 1613 cm⁻¹.

NMR (D₂ O-DCl, δ) 1.30 (9H, s), 3.3 (6H, m), 3.6-4.3 (2H, m), 4.6-5.2(6H, m), 5.60 (1H, d, J=5Hz), 5.90 (1H, d, J=5Hz), 7.33 (1H, s), 7.70(1H, s), 8.30 (1H, s), 8.41 (1H, s).

(3)7β-[2-(2-Formamidothiazol-4-yl)-2-methoxyiminoacetamido]-3-[N,N-dimethyl-N-{2-(5-hydroxy-4-oxo-1,4-dihydropyridin-2-yl)ethyl]ammonio]methyl-3-cephem-4carboxylate(syn isomer)

IR (Nujol):3200 (br), 1772, 1670, 1608 cm⁻¹.

NMR (D₂ O-DCl, δ) 3.23 (3H, s), 3.30 (3H, s), 3.2-4.5 (6H, m), 4.6-5.1(2H, m), 5.43 (1H, d, J=5Hz), 5.83 (1H, d, J=5Hz), 7.58 (1H, s), 7.80(1H, s), 8.07 (1H, s), 8.50 (1H, s).

EXAMPLE 2

(1) To a solution of7β-[2-(5-amino-1,2,4-thiadiazol-3-yl)-2-(1-carboxy-1-methylethoxyimino)acetamido]-3-chloromethyl-3-cephem-4-carboxylicacid trifluoroacetate (syn isomer) (1.0 g) in N,N-dimethylformamide (10ml) was added 2-(N,N-dimethylamino)methyl-5-hydroxy-4pyridone (1.09 g).After being stirred for 5 hours at ambient temperature, the mixture waspoured into ethyl acetate (150 ml). The resulting precipitate wascollected by filtration, washed with ethyl acetate and dried underreduced pressure. The precipitate was suspended in water (50 ml) at pH2.0 and stirred for 30 minutes. After removal of insoluble material, theaqueous solution was subjected to column chromatography on Diaion HP-20.The column was washed with water and the elution was carried out with30% aqueous methanol. The fractions containing desired product wascombined and methanol was evaporated in vacuo. The resulting aqueouslayer was lyophilized to give7β-[2-(5-amino-1,2,4-thiadiazol-3-yl)-2-(1-carboxy-1-methylethoxyimino)acetamido]-3-[N,N-dimethyl-N-{(5-hydroxy-4-oxo-1,4-dihydropyridin-2-yl)methyl}ammonio]methyl-3-cephem-4carboxylate(syn isomer) (0.30 g) as a powder.

IR (Nujol) : 3250 (br), 1770, 1670 (sh), 1600 cm⁻¹.

NMR (D₂ O-NaHCO₃, δ) 1.56 (6H, s), 3.03 (3H, br. s), 3.10 (3H, br s),3.44, 3.96 (2H, ABq, J=18Hz), 4.1-5.0 (4H, m), 5.37 (1H, d, J=5Hz), 5.88(1H, d, J=5Hz), 6.76 (1H, s), 7.71 (1H, s).

The following compounds were obtained according to a similar manner tothat of Example 2(1).

(2)7β-[2-(5-Amino-1,2,4-thiadiazol-3-yl)-2-methoxyiminoacetamido]-3-[1-{2-(5-hydroxy-4-oxo-1,4-dihydropyridin-2-yl)ethyl}-1-pyrrolidinio]methyl-3-cephem-4-carboxylate(syn isomer)

IR (Nujol):3300 (br), 1765, 1663 (sh), 1620 cm⁻¹.

NMR (D₂ O-NaHCO₃, δ) 2.05 (4H, m), 3.0-3.8 (10H, m), 4.07 (3H, s),4.6-4.9 (2H, m), 5.18 (1H, d, J=5Hz), 5.80 (1H, d, J=5Hz), 6.45 (1H, s),7.51 (1H, s).

(3)7β-[2-(5-Amino-1,2,4-thiadiazol-3-yl)-2-(1-carboxy-1-methylethoxyimino)acetamido]-3-[1-{2-(5-hydroxy-4-oxo-1,4-dihydropyridin-2-yl)ethyl}-1-pyrrolidinio]-methyl-3-cephem-4-carboxylate(syn isomer)

IR (Nujol):3300 (br), 1770, 1670-1620 (br) cm⁻¹.

NMR (D₂ O-NaHCO₃, δ) 1.53 (6H, s), 2.06 (4H, m), 3.0-3.8 (10H, m),4.4-4.9 (2H, m), 5.10 (1H, d, J=5Hz), 5.80 (1H, d, J=5Hz), 6.43 (1H, s),7.85 (1H, s).

(4)7β-[2-(5-Amino-1,2,4-thiadiazol-3-yl)-2-(1-carboxy-1-methylethoxyimino)acetamido]-3-[N,N-dimethyl-N-{2-(5-hydroxy-4-oxo-1,4-dihydropyridin-2-yl)ethyl}ammonio]methyl-3-cephem-4-carboxylate(syn isomer)

IR (Nujol):3300 (br), 1775, 1670, 1615 cm⁻¹. NMR (D₂ O-NaHCO₃, δ) 1.55(6H, s), 3.00 (3H, br.s), 3.15 (3H, br. s), 3.2-4.2 (4H, m), 4.6-4.9(2H, m), 5.35 (1H, d, J=5Hz), 5.80 (1H, d, J=5Hz), 6.45 (1H, s), 7.54(1H, s).

(5)7β-[2-(5-Amino-1,2,4-thiadiazol-3-yl)-2-methoxyiminoacetamido]-3-[N,N-dimethyl-N-{2-(5-hydroxy-4-oxo-1,4-dihydropyridin-2-yl)ethyl}ammonio]methyl-3-cephem-4-carboxylate (syn isomer)

IR (Nujol):3250 (br), 1770, 1670(sh), 1610 cm⁻¹.

NMR (D₂ O-NaHCO₃, δ) 3.06 (3H, br. s), 3.13 (3H, br.s), 3.2-4.2 (4H, m),4.03 (3H, s), 4.6-4.9 (2H, m), 5.31 (1H, d, J=5Hz), 5.80 (1H, d, J=5Hz),6.45 (1H, s), 7.50 (1H, s).

(6)7β-[2-(2-Formamidothiazol-4-yl)-2-(1-tert-butoxycarbonyl-1-methylethoxyimino)acetamido]-3-[N,N-dimethyl-N-{2-(5-hydroxy-4-oxo-1,4-dihydropyridin-2-yl)ethyl}ammonio]methyl-3-cephem-4-carboxylate(syn isomer)

IR (Nujol):3200, 1770, 1720, 1685 (sh), 1668, 1605 cm⁻¹.

NMR (D₂ O-NaHCO₃, δ) 1.45 (9H, s), 1.55 (6H, s), 2.10 (3H, s), 3.16 (3H,s), 3.0-4.6 (6H, m), 4.5-4.9 (2H, m), 5.37 (1H, d, J=5Hz), 5.85 (1H, d,J=5Hz), 6.47 (1H, s), 7.45 (1H, s), 7.54 (1H, s), 8.50 (1H, s) .

(7)7β-[2-(2-Formamidothiazol-4-yl)-2-(1-tert-butoxycarbonyl-1-methylethoxyimino)acetamido]-3-[N,N-dimethyl-N-{(5-hydroxy-4-oxo-1,4-dihydropyridin-2-yl)methyl}ammonio]methyl-3-cephem-4-carboxylate(syn isomer)

IR (Nujol):3250, 1780, 1720, 1675, 1611 cm⁻¹.

NMR (D₂ O-DCl, δ) 1.35 (9H, s), 1.60 (6H, s), 3.30 (3H, s), 3.37 (3H,s), 3.6-4.6 (4H, m), 4.92 (2H, br.s), 5.53 (1H, d, J=5Hz), 5.90 (1H, d,J=5Hz), 7.28 (1H, s), 7.70 (1H, s), 8.30 (1H, s), 8.37 (1H, s).

(8)7β-[2-(2-Aminothiazol-4-yl)-2-tert-butoxycarbonylmethoxyiminoacetamido]-3-[N,N-dimethyl-N-{(5-hydroxy-4-oxo-1,4-dihydropyridin-2-yl)methyl}ammonio]methyl-3-cephem-4-carboxylate(syn isomer)

IR (Nujol):3300 (br.), 1750, 1730 (sh), 1667, 1608 cm⁻¹.

(9)7β-[2-(2-Aminothiazol-4-yl)-2-methoxyiminoacetamido]-3-[N,N-dimethyl-N-{2-(5-hydroxy-4-oxo-1,4-dihydropyridin-2-yl)ethyl}ammonio]methyl-3-cephem-4-carboxylate(syn isomer)

IR (Nujol) : 3250 (br), 1770, 1660, 1705 cm⁻¹.

(10)7β-[2-(2-Aminothiazol-4-yl)-2-(1-tert-butoxycarbonyl-1-methylethoxyimino)acetamido]-3-[N,N-dimethyl-N-(2-(5-hydroxy-4-oxo-1,4-dihydropyridin-2-yl)ethyl}ammonio]methyl-3-cephem-4-carboxylate(syn isomer)

IR (Nujol):3250 (br.), 1770, 1720, 1662, 1608 cm⁻¹.

(11)7β-[2-(2-Aminothiazol-4-yl)-2-(1-tert-butoxycarbonyl-1-methylethoxyimino)acetamido]-3-[N,N-dimethyl-N{(5-hydroxy-4-oxo-1,4-dihydropyridin-2-yl)methyl}ammonio]methyl-3-cephem-4-carboxylate(syn isomer)

IR (Nujol):3350 (br.), 1775, 1720, 1672, 1608 cm⁻¹.

(12)7β-[2-(2-Aminothiazol-4-yl)-2-carboxymethoxyiminoacetamido]-3-[N,N-dimethyl-N-{(5-hydroxy-4-oxo-1,4-dihydropyridin-2-yl)methyl}ammonio]methyl-3-cephem-4carboxylate(syn isomer)

IR (Nujol):3250 (br), 1770, 1662 (sh), 1600 cm⁻¹.

(13)7β-[2-(2-Aminothiazol-4-yl)-2-(1-carboxy-1-methylethoxyimino)acetamido]-3-[N,N-dimethyl-N-{2-(5-hydroxy-4-oxo-1,4-dihydropyridin-2-yl)ethyl}ammonio]methyl-3-cephem-4-carboxylate(syn isomer)

IR (Nujol) : 3250 (br), 1770, 1665, 1610 cm⁻¹.

(14)7β-[2-(2-Aminothiazol-4-yl)-2-(1-carboxy-1methylethoxyimino)acetamido]-3-[N,N-dimethyl-N-{(5-hydroxy-4-oxo-1,4-dihydropyridin-2-yl)methyl}ammonio]-methyl-3-cephem-4-carboxylate(syn isomer)

IR (Nujol):3250, 1770, 1662, 1607 cm⁻¹.

(15)7β-[2-(5-Amino-1,2,4-thiadiazol-3-yl)-2-(1-carboxy-1-methylethoxyimino)acetamido]-3-[N,N-dimethyl-N-{(5-hydroxy-4-oxo-1,4-dihydropyridin-2-yl)methyl}ammonio]-methyl-3-cephem-4-carboxylatehydrochloride (syn isomer)

IR (Nujol):3450-3150 (br), 2650, 1770, 1670, 1610 cm⁻¹.

(16) Sulfuric acid salt of7β-[2-(5-amino-1,2,4-thiadiazol-3-yl)-2-(1-carboxy-1-methylethoxyimino)acetamido]-3-[N,N-dimethyl-N-{(5-hydroxy-4-oxo-1,4-dihydropyridin-2-yl)methyl}ammonio]methyl-3-cephem-4carboxylate(syn isomer)

IR (Nujol):3450-3150 (br), 2650 (br), 1780, 1692, 1615, 1558, 1529 cm⁻¹.

EXAMPLE 3

(1) To a suspension of 7β-[2-(2-formamidothiazol-4-yl)-2-tert-butoxycarbonylmethoxyiminoacetamido]-3-[N,N-dimethyl-N-{(5-hydroxy-4-oxo-1,4-dihydropyridin-2-yl)methyl}ammonio]methyl-3-cephem-4-carboxylate(syn isomer) (0.70 g) in methanol (35 ml) was added conc. hydrochloricacid (0.53 ml). After being stirred at ambient temperature, the reactionmixture was diluted with water (50 ml) and methanol was evaporated underreduced pressure. The resulting aqueous solution was adjusted to pH 1.0and subjected to column chromatography on Diaion HP-20 (50 ml). Thecolumn was washed with water and the elution was carried out with 40%aqueous isopropanol. The fractions containing the object compound waslyophilized to give7β-[2-(2-aminothiazol-4-yl)-2-tert-butoxycarbonylmethoxyiminoacetamido]-3-[N,N-dimethyl-N-{(5-hydroxy-4-oxo-1,4-dihydropyridin-2-yl)methyl}-amino]methyl-3-cephem-4-carboxylate (syn isomer) (0.53 g).

IR (Nujol):3300 (br), 1750, 1730 (sh), 1667, 1608 cm⁻¹.

NMR (D₂ O-NaHCO₃, 67 ) 1.50 (9H, s), 2.97 (3H, br.s), 3.06 (3H, br. s),3.2-4.2 (4H, m), 4.36 (2H, br. s), 4.65 (2H, s), 5.35 (1H, d, J=5Hz),5.86 (1H, d, J=5Hz), 6.73 (1H, s), 7.00 (1H, s), 7.73 (1H, s).

The following compounds were obtained according to a similar manner tothat of Example 3(1).

(2)7β-[2-(2-Aminothiazol-4-yl)-2-methoxyiminoacetamido]-3-[N,N-dimethyl-N-{2-(5-hydroxy-4-oxo-1,4-dihydro-pyridin-2-yl)ethyl}ammonio]methyl-3-cephem-4-carboxylate(syn isomer)

IR (Nujol):3250 (br), 1770, 1660, 1705 cm⁻¹.

NMR (D₂ O-NaHCO₃,δ):3.10 (3H, s), 3.15 (3H, s), 3.0-4.2 (6H, m), 4.5-4.9(2H, m), 5.33 (1H, d, J=5Hz), 5.80 (1H, d, J=5Hz), 6.50 (1H, s), 6.95(1H, s), 7.51 (1H, s).

(3)7β-[2-(2-Aminothiazol-4-yl)-2-(1-tert-butoxycarbonyl-1-methylethoxyimino)acetamido]-3-[N,N-dimethyl-N-{2-(5-hydroxy-4-oxo-1,4-dihydropyridin-2-yl)ethyl}ammonio]methyl-3-cephem-4-carboxylate(syn isomer)

IR (Nujol):3250 (br), 1770, 1720, 1662, 1608 cm⁻¹.

NMR (D₂ O-NaHCO₃, δ) 1.50 (15H, s), 2.10 (3H, s), 2.17 (3H, s), 3.0-4.2(6H, m), 4.6-5.0 (2H, m), 5.36 (1H, d, J=5Hz), 5.83 (1H, d, J=5Hz), 6.50(1H, s), 6.93 (1H, s), 7.56 (1H, s).

(4)7β-[2-(2-Aminothiazol-4-yl)-2-(1-tert-butoxycarbonyl-1-methylethoxyimino)acetamido]-3-[N,N-dimethyl-N-}(5-hydroxy-4-oxo-1,4-dihydropyridin-2-yl)methyl}-ammonio]methyl-3-cephem-4-carboxylate(syn isomer)

IR (Nujol):3350 (br), 1775, 1720, 1672, 1608 cm⁻¹.

NMR (D₂ O-NaHCO₃, δ):1.45 (9H, s), 1.55 (6H, s), 2.96 (3H, s), 3.05 (3H,s), 3.25-4.2 (4H, m), 4.36 (2H, br. s), 5.35 (1H, d, J=5Hz), 5.85 (1H,d, J=5Hz), 6.72 (1H, s), 6.95 (1H, s), 7.71 (1H, s).

(5)7β-[2-(5-Amino-1,2,4-thiadiazol-3-yl)-2-methoxyiminoacetamido]-3-[N,N-dimethyl-N-{(5-hydroxy-4-oxo1,4-dihydropyridin-2-yl)methyl}ammonio]methyl-3-cephem-4-carboxylate(syn isomer)

IR (Nujol):3250 (br), 1765, 1650, 1600 cm⁻¹.

(6)7β-[2-(5-Amino-1,2,4-thiadiazol-3-yl)-2-(1-carboxy-1-methylethoxyimino)acetamido]-3-[N,N-dimethyl-N-{(5-hydroxy-4-oxo-1,4-dihydropyridin-2-yl)methyl}ammonio]methyl-3-cephem-4-carboxylate(syn isomer)

IR (Nujol):3250 (br.), 1770, 1670 (sh), 1600 cm⁻¹.

(7)7β-[2-(5-Amino-1,2,4-thiadiazol-3-yl)-2-methoxyiminoacetamido]-3-[1-{2-(5-hydroxy-4-oxo-1,4-dihydropyridin-2-yl)ethyl}-1-pyrrolidinio]methyl-3-cephem-4-carboxylate(syn isomer)

IR (Nujol):3300 (br.), 1765, 1663 (sh), 1620 cm⁻¹.

(8)7β-[2-(5-Amino-1,2,4-thiadiazol-3-yl)-2-(1-carboxy-1-methylethoxyimino)acetamido]-3-[1-{2-(5-hydroxy-4-oxo-1,4-dihydropyridin-2-yl)ethyl}-1pyrrolidinio]methyl-3-cephem-4-carboxylate(syn isomer)

IR (Nujol):3300 (br), 1770, 1670, 1620 (br.) cm⁻¹.

(9)7β-[2-(5-Amino-1,2,4-thiadiazol-3-yl)-2-(1-carboxy-1-methylethoxyimino)acetamido]-3-[N,N-dimethyl-N-{2-(5-hydroxy-4-oxo-1,4-dihydropyridin-2-yl)ethyl}ammonio]-methyl-3-cephem-4-carboxylate(syn isomer)

IR (Nujol):3300 (br.), 1775, 1670, 1615 cm⁻¹.

(10)78-[2-(5-Amino-1,2,4-thiadiazol-3-yl)-2-methoxyiminoacetamido]-3-[N,N-dimethyl-N-{2-(5-hydroxy-4-oxo-1,4-dihydropyridin-2-yl)ethyl}ammonio]methyl-3-cephem-4-carboxylate(syn isomer)

IR (Nujol):3250 (br), 1770, 1670 (sh), 1610 cm⁻¹.

(11)7β-[2-(2-Aminothiazol-4-yl)-2-carboxymethoxyiminoacetamido]-3-[N,N-dimethyl-N-{(5-hydroxy-4-oxo-1,4-dihydropyridin-2-yl)methyl}ammonio]methyl-3-cephem-4-carboxylate(syn isomer)

IR (Nujol):3250 (br.), 1770, 1662 (sh), 1600 cm⁻¹.

(12)7β-[2-(2-Aminothiazol-4-yl)-2-(1-carboxy-1-methylethoxyimino)acetamido]-3-[N,N-dimethyl-N-{2-(5-hydroxy-4-oxo-1,4-dihydropyridin-2-yl)ethyl}ammonio]methyl-3-cephem-4-carboxylate(syn isomer)

IR (Nujol):3250 (br), 1770, 1665, 1610 cm⁻¹.

(13)7β-[2-(2-Aminothiazol-4-yl)-2-(1-carboxy-1-methylethoxyimino)acetamido]-3-[N,N-dimethyl-N-{(5-hydroxy-4-oxo-1,4-dihydropyridin-2-yl)methyl}ammonio]methyl-3-cephem-4-carboxylate(syn isomer)

IR (Nujol):3250, 1770, 1662, 1607 cm⁻¹.

EXAMPLE 4

(1) To a suspension of7β-[2-(2-aminothiazol-4-yl)-2-tert-butoxycarbonylmethoxyiminoacetamido]-3-[N,N-dimethyl-N-{(5-hydroxy-4-oxo-1,4-dihydropyridin-2-yl)methyl}ammonio]methyl-3-cephem-4-carboxylate(syn isomer) (0.54 g) in anisole (2 ml) was dropwise addedtrifluoroacetic acid (8 ml) under ice-cooling with stirring. After beingstirred at ambient temperature, the mixture was poured into diisopropylether (100 ml). The resulting precipitate was collected by filtration,washed with diisopropyl ether and dried under reduced pressure. Theprecipitate was dissolved in water (30 ml) at pH 7.0 and acidified to pH1.0 with 6N hydrochloric acid. After removal of precipitated materials,the aqueous solution was subjected to column chromatography on DiaionHP-20 (50 ml). The column was washed with water and the elution wascarried out with 30% aqueous methanol. The active fractions werecollected and lyophilized to give7β-[2-(2-aminothiazol-4-yl)-2-carboxymethoxyiminoacetamido]-3-[N,N-dimethyl-N-{(5-hydroxy-4-oxo-1,4-dihydropyridin-2-yl)methyl}ammonio]-methyl-3-cephem-4-carboxylate(syn isomer) (0.28 g).

IR (Nujol):3250 (br), 1770, 1662 (sh), 1600 cm⁻¹.

NMR (D₂ O-NaHCO₃, δ):3.00 (3H, s), 3.07 (3H, s), 3.40, 3.94 (2H, ABq,J=18Hz), 4.1-5.1 (4H, m), 4.45 (2H, br. s), 4.60 (2H, s), 5.35 (1H, d,J=5Hz), 5.86 (1H, d, J=5Hz), 6.75 (1H, s), 6.93 (1H, s), 7.70 (1H, s) .

The following compounds were obtained according to a similar manner tothat of Example 4(1).

(2)7β-[2-(2-Aminothiazol-4-yl)-2-(1-carboxy-1-methylethoxyimino)acetamido]-3-[N,N-dimethyl-N-{2-(5-hydroxy-4-oxo-1,4-dihydropyridin-2-yl)ethyl}ammonio]-methyl-3-cephem-4-carboxylate(syn isomer)

IR (Nujol):3250 (br), 1770, 1665, 1610 cm⁻¹.

NMR (D₂ O-NaHCO₃, δ):1.50 (6H, s), 3.10 (3H, s), 3.17 (3H, s), 3.0-4.2(6H, m), 4.6-5.0 (2H, m), 5.35 (1H, d, J=5Hz), 5.81 (1H, d, J=5Hz), 6.50(1H, s), 6.91 (1H, s), 7.55 (1H, s).

(3)7β-[2-(2-Aminothiazol-4-yl)-2-(1-carboxy-1-methylethoxyimino)acetamido]-3-[N,N-dimethyl-N-{(5-hydroxy-4-oxo-1,4-dihydropyridin-2-yl)methyl}ammonio]-methyl-3-cephem-4-carboxylate(syn isomer)

IR (Nujol):3250, 1770, 1662, 1607 cm⁻¹.

NMR (D₂ O-NaHCO₃, δ):1.50 (8H, s), 2.98 (3H, s), 3.70 (3H, s), 3.25-4.25(4H, m), 4.42 (2H, br. s), 5.34 (1H, d, J=5Hz), 5.83 (1H, d, J=5Hz),6.74 (1H, s), 6.90 (1H, s), 7.72 (1H, s).

(4)7β-[2-(5-Amino-1,2,4-thiadiazol-3-yl)-2-(1-carboxy-1-methylethoxyimino)acetamido]-3-[N,N-dimethyl-N-{(5-hydroxy-4-oxo-1,4-dihydropyridin-2-yl)methyl}ammonio]-methyl-3-cephem-4-carboxylate(syn isomer)

IR (Nujol):3250 (br), 1770, 1670 (sh), 1600 cm⁻¹.

(5)7β-[2-(5-Amino-1,2,4-thiadiazol-3-yl)-2-(1-carboxy-1-methylethoxyimino)acetamido]-3-[1-{2-(5-hydroxy-4-oxo-1,4-dihydropyridin-2-yl)ethyl}-1pyrrolidinio]methyl-3-cephem-4-carboxylate(syn isomer)

IR (Nujol):3300 (br.), 1770, 1670-1620 (br).

(6)7β-[2-(5-Amino-1,2,4-thiadiazol-3-yl)-2-(1-carboxy-1-methylethoxyimino)acetamido]-3-[N,N-dimethyl-N-{2-(5-hydroxy-4-oxo-1,4-dihydropyridin-2-yl)ethyl}ammonio]methyl-3-cephem-4-carboxylate(syn isomer)

IR (Nujol):3300 (br.), 1775, 1670, 1615 cm⁻¹.

Preparation 6

To a suspension of 5-benzyloxy-2-hydroxymethyl-4-pyridone (33 g) inbenzene (500 ml) was added thionyl chloride (28.4 ml) at ambienttemperature under stirring. After being stirred at the same temperaturefor 30 minutes, the mixture was refluxed for 4 hours. The resultingmixture was cooled to ambient temperature. The precipitate was collectedby filtration, washed with benzene and dried under reduced pressure togive 5-benzyloxy-2-chloromethyl-4-pyridone (41.5 g).

IR (Nujol):1608, 1585 cm⁻¹.

NMR DMSO-d₆, δ):5.00 (2H, s), 5.30 (2H, s), 7.4 (5H, m), 7.56 (1H, s),8.43 (1H, s).

Preparation 7

A suspension of benzhydryl7β-amino-3-chloromethyl-3-cephem-4-carboxylate hydrochloride (36.11 g)in a mixture of ethyl acetate (900 ml) and cold water (360 ml) wasadjusted to pH 7 with a saturated aqueous solution of potassiumcarbonate. The separated organic layer was washed with brine, dried overmagnesium sulfate and evaporated in vacuo. The residue was pulverizedwith diisopropyl ether to give the powder (34.40 g). The powder (34.40g) was added portionwise at 5° C. to acetic formic anhydride preparedfrom formic acid (15.27 g) and acetic anhydride (33.89 g). The mixturewas warmed to room temperature and stirred for 1.8 hours at the sametemperature. The mixture was poured into a mixture of ethyl acetate (1l) and cold water (400 ml) and adjusted to pH 7 with 20% aqueous sodiumhydroxide solution under cooling. The separated organic layer was washedwith water and brine, dried over magnesium sulfate and concentrated to100 ml. The residual solution was poured into a mixture of diisopropylether (1 l) and hexane (1 l) and the resulting precipitates werecollected by filtration to give benzhydryl7β-formamido-3-chloromethyl-3-cephem-4-carboxylate (29.07 g).

IR (Nujol):1780, 1720, 1675 cm⁻¹.

NMR (DMSO-d₆, δ):3.55 and 3.79 (2H, ABq, J=18Hz), 4.53 (2H, s), 5.23(1H, d, J=5Hz), 5.89 (1H, dd, J=8 and 5Hz), 6.96 (1H, s), 7.2-7.6 (10H,m), 8.15 (1H, s), 9.10 (1H, d, J=8Hz).

Preparation 8

To a solution of benzhydryl78-formamido-3-chloromethyl-3-cephem-4-carboxylate (27.34 g) in amixture of dichloromethane (137 ml) and anisole (27 ml) was addeddropwise trifluoroacetic acid (54 ml) and the mixture was stirred at 5°C. for 1.2 hours. The mixture was added dropwise to a cooled mixture ofdiisopropyl ether (2 l) and hexane (2 l) and the resulting precipitateswere collected by filtration, washed with a mixture of diisopropyl etherand hexane to give 7β-formamido-3-chloromethyl-3-cephem-4-carboxylicacid (14.03 g).

IR (Nujol):1775, 1665, 1520 cm⁻¹.

NMR (DMSO-d₆, δ):3.49 and 3.74 (2H, ABq, J=18Hz), 4.55 (2H, s), 5.15(1H, d, J=5Hz), 5.78 (1H, dd, J=8 and 5Hz), 8.12 (1H, s), 9.06 (1H, d,J=8Hz).

Preparation 9

2-(N,N-Dimethylamino)methyl-5-hydroxy-4-pyridone (15.85 g) was dissolvedin N,N-dimethylformamide (238 ml) by addition of sodium 2-ethylhexanoate(10.44 g). To the resulting solution was added dropwise a cooledsolution of 78-formamido-3-chloromethyl-3-cephem-4carboxylic acid (13.04g) at 5° C. and the mixture was stirred for 2 hours at the sametemperature. The mixture was added dropwise to a mixture of ethylacetate (2.5 l) and diisopropyl ether (2.5 l) and the resultingprecipitates were collected by filtration, washed three times with amixture of ethyl acetate and diisopropyl ether (1:1) and dried in vacuo.The powder was poured into cold water (300 ml) and the mixture wasadjusted to pH 3.0 with 1N hydrochloric acid. After the insolublematerials were filtered off, the filtrate was chromatographed on DiaionHP-20 (1300 ml) at 50° C. and the elution was carried out with 10%aqueous isopropyl alcohol. The eluate was lyophilized to give 7β-formamido-3-[N,N-dimethyl-N-{(5-hydroxy-4-oxo-1,4-dihydropyridin-2-yl)methyl}ammonio]methyl-3-cephem-4carboxylate(8.31 g).

IR (Nujol):1775, 1670, 1610, 1565, 1515 cm⁻¹.

NMR (D₂ O, δ):3.05 and 3.14 (6H, s x 2), 3.47 and 3.99 (2H, ABq,J=18Hz), 4.45-4.55 (2H, m), 4.40 and 4.90 (2H, ABq, J=14Hz), 5.31 (1H,d, J=5Hz), 5.80 (1H, d, J=5Hz), 6.84 (1H, s), 7.81 (1H, s), 8.24 (1H,s).

Preparation 10

To a cooled suspension of78-formamido-3-[N,N-dimethyl-N-{(5-hydroxy-4-oxo-1,4-dihydropyridin-2-yl)methyl}ammonio]methyl-3-cephem-4-carboxylate(1.225 g) in methanol (12 ml) was added dropwise conc. hydrochloric acid(0.83 ml) at 10° C. The mixture was warmed to room temperature andstirred for 2.5 hours. The mixture was poured into ethyl acetate (300ml) and the precipitates were collected by filtration, washed with ethylacetate and diisopropyl ether and dried in vacuo. The powder wasdissolved in cold water (10 ml) and the resulting solution waschromatographed on Diaion HP-20 (10 ml) at 5° C. and the elution wascarried out with cold water. To the eluate (24 ml) was added dropwiseisopropyl alcohol (12 ml) under cooling and the mixture was allowed tostand overnight in a refrigerator. The resultant crystal was collectedby filtration, washed with a cold mixture of isopropyl alcohol and water(10:1) and cold isopropyl alcohol, and dried in vacuo to give 7β-amino-3-[N,N-dimethyl-N-{(5-hydroxy-4-oxo-1,4-dihydropyridin-2-yl)methyl}ammonio]methyl-3-cephem-4-carboxylatedihydrochloride (440 mg).

IR (Nujol):3350, 1810, 1790, 1640, 1520 cm⁻¹.

NMR (D₂ O+NaHCO₃, δ):3.04 and 3.14 (6H, s×2), 3.48 and 3.99 (2H, ABq,J=18Hz), 4.13 and 4.72 (2H, ABq, J=14Hz), 4.45-4.55 (2H, m), 4.94 (1H,d, J=5Hz), 5.29 (1H, d, J=5Hz), 6.85 (1H, s), 7.82 (1H, s).

Preparation 11

To a cooled solution of7β-formamido-3-[N,N-dimethyl-N-{(5-hydroxy-4-oxo-1,4-dihydropyridin-2-yl)methyl}ammonio]methyl-3-cephem-4-carboxylate(4.084 g) in formic acid (8.1 ml) was added dropwise conc. hydrochloricacid (2.5 ml). The mixture was warmed to room temperature and stirredfor 2.5 hours. The mixture was added dropwise to ethyl acetate (400 ml)and the supernatant was decanted. The residual oil was dissolved inmethanol (50 ml) and the solution was poured into ethyl acetate (600ml). The resulting precipitates were collected by filtration, washedwith ethyl acetate and diisopropyl ether and dried in vacuo. The powderwas dissolved in cold water (28 ml) and chromatographed on Diaion HP-20(28 ml) at 5° C., and the elution was carried out with water. To theeluate (30 ml) was added dropwise cold isopropyl alcohol (19 ml) undercooling and the mixture was stirred at 5° C. for an hour. The resultantcrystal was collected by filtration washed with a cooled mixture ofisopropyl alcohol and water (10:1) and cold isopropyl alcohol, and driedin vacuo to give7β-amino-3-[N,N-dimethyl-N-{(5-hydroxy-4-oxo-1,4-dihydropyridin-2-yl)methyl}ammonio]methyl-3-cephem-4carboxylatedihydrochloride (728 mg).

IR (Nujol):3350, 1810, 1790, 1640, 1520 cm⁻¹.

NMR (D₂ O+NaHCO₃, δ):3.04 and 3.14 (6H, s×2), 3.48 and 3.99 (2H, ABq,J=18Hz), 4.13 and 4.72 (2H, ABq, J=14Hz), 4.45-4.55 (2H, m), 4.94 (1H,d, J=5Hz), 5.29 (1H, d, J=5Hz), 6.85 (1H, s), 7.82 (1H, s).

EXAMPLE 5

7β-amino-3-[N,N-dimethyl-N-{(5-hydroxy-4-oxo-1,4-dihydropyridin-2-yl)methyl}ammonio]methyl-3-cephem-4-carboxylatedihydrochloride (181 mg) was suspended in a mixture of water (7.2 ml)and tetrahydrofuran (3.6 ml) and adjusted to pH 5 with a saturatedaqueous solution of sodium bicarbonate. To the resulting solution wasadded portionwise a solution of2-(5-amino-1,2,4-thiadiazol-3-yl)-2-(1-carboxy-1-methylethoxyimino)aceticmethanesulfonic anhydride (syn isomer) (169 mg) in tetrahydrofuran (0.5ml) at 15° C. and the mixture was stirred for an hour at 15° C. and pH4.0-6.0. After tetrahydrofuran was evaporated in vacuo, water (30 ml)was added to the residue. The aqueous solution was adjusted to pH 1.0with 1N hydrochloric acid and filtered to remove the insolublematerials. The filtrate was chromatographed on Diaion HP-20 (15 ml) andthe elution was carried out with 15% aqueous isopropyl alcohol. Theeluate was lyophilized to give

7β-[2-(5-amino-1,2,4-thiadiazol-3-yl)-2-(1-carboxy-1methylethoxyimino)acetamido]-3-[N,N-dimethyl-N-{(5-hydroxy-4-oxo-1,4-dihydropyridin-2-yl)methyl}ammonio]-methyl-3-cephem-4-carboxylate(syn isomer) (124 mg).

IR (Nujol):3250 (br), 1770, 1670 (sh), 1600 cm⁻¹.

NMR (D₂ O-NaHCO₃, δ) 1.56 (6H, s , 3.03 (3H, br s), 3.10 (3H, br s),3.44, 3.96 (2H, ABq, J=18Hz), 4.1-5.0 (4H, m), 5.37 (1H, d, J=5Hz), 5.88(1H, d, J=5Hz), 6.76 (1H, s), 7.71 (1H, s).

EXAMPLE 6

7β-Amino-3-[N,N-dimethyl-N-{(5-hydroxy-4-oxo-1,4-dihydropyridin-2-yl)methyl}ammonio]methyl-3-cephem-4-carboxylatedihydrochloride (181 mg) was suspended in tetrahydrofuran (9.1 ml). Tothe suspension was added N-(trimethylsilyl)acetamide (788 mg), and themixture was stirred at 35° C. for 30 minutes and cooled to 5° C. To theresulting solution was added a solution of2-(5-amino-1,2,4-thiadiazol-3-yl)-2-(1-carboxy-1-methylethoxyimino)aceticmethanesulfonic anhydride (syn isomer) (169 mg) in tetrahydrofuran (0.5ml). The mixture was warmed to 20° C. and stirred for 1.2 hours. Themixture was poured into ethyl acetate (200 ml) and the resultingprecipitates were collected by filtration, washed with ethyl acetate anddiisopropyl ether and dried in vacuo. The powder was suspended in water(40 ml) and adjusted to pH 1 with 1N hydrochloric acid. After theinsoluble materials were removed by filtration, the filtrate waschromatographed on Diaion HP-20 (15 ml) and the elution was carried outwith 15% aqueous isopropyl alcohol. The eluate was lyophilized to give7β-[2-(5-amino-1,2,4-thiadiazol-3-yl)-2-(1-carboxy-1-methylethoxyimino)acetamido]-3-[N,N-dimethyl-N-{(5-hydroxy-4-oxo-1,4-dihydropyridin-2-yl)methylammonio]methyl-3-cephem-4-carboxylate(syn isomer) (120 mg).

IR (Nujol):3250 (br), 1770, 1670 (sh), 1600 cm⁻¹.

Preparation 12

A mixture of 4,5-dihydroxy-2-pyridinecarboxylic acid dihydrate (15.5 g)in acetic anhydride (77.5 ml) was heated at 90°-95° C. for 30 minutes.After the mixture was cooled, the solvent was evaporated in vacuo. Tothe residue were added acetone (20 ml) and water (10 ml) and the mixturewas stirred for 1.5 hours at room temperature. The mixture wasevaporated in vacuo to remove acetone. The aqueous residue was extractedwith ethyl acetate and the extract was washed with a saturated aqueoussolution of sodium chloride, dried over magnesium sulfate, evaporated invacuo and triturated with diethyl ether to give4,5-diacetoxy-2-pyridinecarboxylic acid (14.27 g).

IR (Nujol):1780, 1750, 1700, 1590 cm⁻¹.

NMR (DMSO-d₆, δ):2.43 (6H, s), 8.15 (1H, s), 8.75 (1H, s).

Preparation 13

To a suspension of phosphorus pentachloride (4.6 g) in methylenechloride (40 ml) was added 4,5-diacetoxy-2-pyridinecarboxylic acid (4.5g) at -20° C. with stirring. The stirring was continued for an hour at-18°˜12° C. To the reaction mixture was added diisopropyl ether (80 ml)below 0° C. The resulting precipitates were collected by filtration,washed with diisopropyl ether and dried over phosphorus pentoxide togive 4,5-diacetoxy-2-pyridinecarbonyl chloride hydrochloride (5.15 g).

IR (Nujol):1790, 1750, 1615 cm⁻¹.

Preparation 14

To a solution of N-methylpiperazine (300 mg) in methylene chloride (10ml) was added dropwise a solution of 3,4-diacetoxybenzoyl chloride (924mg) in methylene chloride (4 ml) at 0° C., and then triethylamine (364mg) was added to the mixture.

The mixture was stirred at 0° C. for 20 minutes and at room temperaturefor 1 hour. Ice water (10 ml) was added to the mixture and the organiclayer was separated, washed with water and brine, dried over magnesiumsulfate and evaporated in vacuo. The residue was purified by columnchromatography on silica gel (20 g) [eluent:choloroform-methanol (25:1)]to give 1-methyl-4-(3,4-diacetoxybenzoyl)piperazine (470 mg) as viscousoil.

IR (Nujol): 2950, 2810, 1765, 1630, 1505 cm⁻¹.

NMR (DMSO-d₆, δ):2.16 (3H, s), 2.26 (6H, s), 2.1-2.4 (4H, m), 3.1-3.6(4H, m), 7.31 (3H,s).

Preparation 15

To a suspension of 2-methoxycarbonyl-4-hydroxy-5-benzyloxypyridine(18.15 g) in N-methylpiperazine (21.04 g) was heated to 100° C. for 3.5hours. The resulting viscous solution was cooled to room temperature,diluted with a mixture of chloroform (270 ml) and methanol (30 ml) andsubjected to column chromatography on silica gel (800 g). The eluate wasevaporated in vacuo and the residue was purified by columnchromatography on silica gel (600 g). The pure fractions were evaporatedin vacuo and the residue was pulverized with diisopropyl ether to give1-methyl-4-(4-hydroxy-5-benzyloxy-2-pyridylcarbonyl)piperazine (3.99 g).

IR (Nujol):1645, 1610, 1540, 1505 cm⁻¹.

NMR (DMSO-d₆, δ):2.18 (3H, s), 2.30 (4H, m), 3.51 (4H, m), 5.16 (2H, s),6.88 (1H, s), 7.3-7.6 (5H, m), 8.02 (1H, s).

Preparation 16

To a solution of 1-methyl-4-(3,4-diacetoxybenzoyl)piperazine (295 mg) nmethanol (3 ml) was added a saturated potassium carbonate solution ismethanol (6 ml) and the mixture was stirred at room temperature for 30minutes. The mixture was poured into ice water and adjusted to pH 9 with1N hydrochloric acid. The mixture was extracted several times with amixture of chloroform and methanol (20:1 V/V) and the organic layer waswashed with brine, dried over magnesium sulfate and evaporated in vacuo.The residue was crystallized by addition of ethyl acetate to give1-methyl-4-(3,4-dihydroxybenzoyl)piperazine (71 mg) as white crystal.

mp:222°-225° C.

IR (Nujol):3160, 1590, 1530 cm⁻¹.

NMR (DMSO-d₆, δ):2.20 (3H, s), 2.2-2.4 (4H, m), 3.3-3.6 (4H, m), 6.6-6.8(3H, m), 8.7-9.4 (2H, br).

Preparation 17

To a solution of1-methyl-4-(4-hydroxy-5-benzyloxy-2-pyridylcarbonyl)piperazine (4.71 g)in methanol (300 ml) was added 10 palladium-carbon (2.35 g) in a streamof nitrogen and the mixture was hydrogenated at atmospheric pressure for20 minutes. After the catalyst was removed by filtration, the filtratewas evaporated to dryness. The residue was recyrstallized from methanolto give 1-methyl-4-(4,5-dihydroxy-2-pyridylcarbonyl)piperazine (1.32 g).

mp:120°-123° C.

IR (Nujol):1630, 1545 cm⁻¹.

NMR (D₂ O, δ):2.51 (3H, s), 2.85 (4H, m), 3.72 (4H, m), 6.67 (1H, s),7.71 (1H, s).

Preparation 18

To a solution of benzhydryl7β-amino-3-chloromethyl-3-cephem-4-carboxylate hydrochloride (22.6 g)and N-trimethylsilylacetamide (25.0 g) in tetrahydrofuran (250 ml) wasadded 2-(5-amino-1,2,4-thiadiazol-3-yl)-2-carboxymethoxyiminoaceticmethanesulfonic anhydride (syn isomer) (19.8 g) at 0°-5° C. understirring. The stirring was continued for an hour at the sametemperature. The reaction mixture was poured into a mixture of ethylacetate (500 ml) and water (500 ml). The organic layer was separated,washed with water twice and a saturated aqueous solution of sodiumchloride, dried over magnesium sulfate and then concentrated in vacuo.To the residue was added diethyl ether and then the resultingprecipitates were collected by filtration to give benzyhydryl7β-[2-(5-amino-1,2,4-thiadiazol-3-yl)-2-carboxymethyoxyiminoacetamido]-3-chloromethyl-3-cephem-4-carboxylate(syn isomer) (17.2 g).

NMR (DMSO-d₆, δ):3.57, 3.70 (2H, Abq, J=18Hz), 4.42 (2H, s), 4.63 (2H,s), 5.22 (1H, d, J=5Hz), 5.90 (1H, dd, J=5Hz, J=8Hz), 6.92 (1H, s),7.17-7.53 (5H, m), 8.07 (2H, br s), 9.52 (1H, d, J=8Hz).

Preparation 19

To a solution of trifluoroacetic acid (20 ml) and anisole (10 ml) inmethylene chloride (50 ml) was added benzylhydryl7β-[2-(5-amino-1,2,4-thiadiazol-3-yl)-2-carboxymethoxyimioacetamido]-3-chloromethyl-3-cephem-4-carboxylate(syn isomer) (7.0 g) at 0˜-5° C. under stirring. The stirring wascontinued for 2 hours at the same temperature. The reaction mixture waspoured into a cold mixture of diisopropyl ether (400 ml) and n-hexane(200 ml). The resulting precipitates were collected by filtration togive7β-[2-(5-amino-1,2,4-thiadiazol-3-yl)-2-carboxymethoxyiminoacetamido]-3-chloromethyl-3-cephem-4-carboxylicacid (syn isomer) (5.5 g).

IR (Nujol):3300, 1770, 1700, 1670, 1610, 1510 cm⁻¹.

NMR (DMSO-d₆, δ):3.53, 3.73 (2H, ABq, J=18Hz), 4.55 (2H, s), 4.65 (2H,s), 5.17 (1H, d, J=5Hz), 5.83 (1H, dd, J=5Hz, J=8Hz), 8.10 (2H, br s),9.48 (1H, d, J=8Hz).

Preparation 20

(1) To a solution of78-[2-(5-amino-1,2,4-thiadiazol-3-yl)-2-carboxymethoxyiminoacetamido]-3-chloromethyl-3-cephem-4-carboxylicacid (syn isomer) (4.0 g) in N,N-dimethylformamide was added dropwise1-methyl-4-tert-butoxycarbonylpiperazine (6.7 g) at 20° C. understirring. The stirring was continued for 1.5 hours at 18°-20° C. Thereaction mixture was poured into ethyl acetate (200 ml). The resultingprecipitates were collected by filtration to give78-[2-(5-amino-1,2,4-thiadiazol-3-yl)-2-carboxymethoxyiminoacetamido]-3-(1-methyl-4-tert-butoxycarbonyl-1-piperazinio)methyl-3-cephem-4-carboxylatehydrochloride (syn isomer) (5.71 g).

The following compounds were obtained according to a similar manner tothat of Preparation 20(1).

(2)7β-[2-(5-amino-1,2,4-thiadiazol-3-yl)-2-ethoxyiminoacetamido]-3-(1-methyl-4-tert-butoxycarbonyl-1-piperazinio)methyl-3-cephem-4-carboxylatehydrochloride trifluoroacetate (syn isomer)

IR (Nujol):1770, 1660, 1610, 1520 cm⁻¹.

NMR (DMSO-d6, 6):1.26 (3H, t, J=7Hz), 2.9-3.1 (7H, m), 3.3-3.9 (6H, m),4.10 (2H, q, J=7Hz), 4.8-5.0 (2H, m), 5.18 (1H, d, J=5Hz), 5.76 (1H, dd,J=8 and 5Hz), 8.16 (2H, br s), 9.49 (1H, d, J=8Hz).

(3)7β-[2-(2-Formamidothiazol-4-yl)-2-tert-butoxycarbonylmethoxyiminoacetamido]-3-(1-methyl-4-tert-butoxycarbonyl-1-piperazinio)methyl-3-cephem-4-carboxylatehydrochloride (syn isomer)

IR (Nujol):1780, 1675, 1610, 1540 cm⁻¹.

NMR (DMSO-d₆, δ):1.43 (18H, s), 3.03 3H, br s), 3.2-4.3 (10H, m), 4.58(2H, s), 4.9-5.1 (2H, m), 5.17 (1H, d, J=5Hz), 5.73 (1H, dd, J=8 andJ=5Hz), 7.37 (1H, s), 8.47 (1H, s), 9.51 (1H, d, J=8Hz).

Preparation 21

(1) To a mixture of formic acid (10 ml) and trifluoroacetic acid (5 ml)was added7β-[2-(5-amino-1,2,4-thiadiazol-3-yl)-2-carboxymethoxyiminoacetamido]-3-(1-methyl-4-tert-butoxycarbonyl-1-piperazinio)methyl-3-cephem-4-carboxylatehydrochloride (syn isomer) (5.7 g) at 20° C. under stirring. Thestirring was continued for 1.5 hours at the same temperature. Thereaction mixture was poured into ethyl acetate (200 ml). The resultingprecipietes were collected by filtration, added to water. The mixturewas adjusted to pH 2.0 with a saturated aqueous solution of sodiumbicarbonate and filtered. The filtrate was subjected to columnchromatography on Diaion HP-20 using 10% aqueous isopropanol as aneluent. The eluate was evaporated in vacuo to remove isopropanol andthen lyophilized to give7β-[2-(5-amino-1,2,4-thiadiazol-3-yl)-2-carboxymethoxyiminoacetamido]-3-(1-methyl-1-piperazinio)methyl-3-cephem-4-carboxylate(syn isomer) (1.50 g).

NMR (D₂ O, δ):3.20 (3H, s), 5.23 (1H, d, J=5Hz), 5.80 (1H, d, J=5Hz).

The following compound was obtained according to a similar manner tothat of Preparation 21(1).

(2)7β-[2-(5-Amino-1,2,4-thiadiazol-3-yl)-2-ethoxyiminoacetamido]-3-(1-methyl-1-piperazinio)methyl-3-cephem-4-carboxylate(syn isomer)

IR (Nujol):1765, 1655, 1600, 1510 cm⁻¹.

NMR (D20, 6):1.33 (3H, t, J=7Hz), 3.20 (3H, br s), 3.3-4.8 (6H, m), 4.34(2H, q, J=7Hz), 4.8-5.0 (2H, m), 5.33 (1H, d, J=5Hz), 5.84 (1H, d,J=5Hz).

Preparation 22

(1) To a solution of7β-[2-(5-amino-1,2,4-thiadiazol-3-yl)-2-methoxyiminoacetamido]-3-chloromethyl-3-cephem-4-carboxylicacid trifluoroacetate (syn isomer) in N,N-dimethylformamide (150 ml) wasadded 1-methyl-4-tert-butoxycarbonylpiperazine (30 g). After beingstirred at room temperature for 30 minutes, the mixture was addeddropwise to ethyl acetate (1.5 l). The resulting precipitate wascollected by filtration, washed with ethyl acetate and diisopropylether. To a solution of the above precipitate in a mixture of methylenechloride (6e ml) and anisole (20 ml) was added dropwise trifluoroaceticacid (40 ml) under ice-cooling. The reaction mixture was stirred at roomtemperature for 2 hours and added to diisopropyl ether (1.5 l) understirring. The resultant precipitate was collected by filtration anddissolved in aqueous sodium bicarbonate, and the solution was adjustedto pH 4.0 with hydrochloric acid and subjected to column chromatographyon "Diaion HP-20" (Trademark:prepared by Mitsubishi Chemical Industries)with 5% aqueous isopropyl alcohol as an eluent. The eluate waslyophilized to give7β-[2-(5-amino-1,2,4-thiadiazol-3-yl)-2-methoxyiminoacetamido]-3-(1-methyl-1-piperazinio)methyl-3-cephem-4-carboxylate(syn isomer) (12.00 g).

IR (Nujol):3300, 1770, 1670, 1610, 1520 cm⁻¹.

The following compound was obtained according to a similar manner tothat of Preparation 22(1).

(2)7β-[2-(2-Formamidothiazol-4-yl)-2-methoxyiminoacetamido]-3-(1-methyl-1-piperazinio)methyl-3-cephem-4-carboxylate(syn isomer)

NMR (D₂ O, δ):3.12 (3H, s), 3.16-4.30 (10H), 4.03 (3H, s), 5.36 (1H, d,J=5Hz), 5.86 (1H, d, J=5Hz), 7.46 (1H, s), 8.43 (1H, s).

Preparation 23

To a suspension of7β-[2-(2-formamidothiazol-4-yl)-2-tert-butoxycarbonylmethoxyiminoacetamido]-3-(1-methyl-4-tert-butoxycarbonyl-1-piperazinio)methyl-3-cephem-4-carboxylatehydrochloride (syn isomer) (4.02 g) in methanol (12 ml) was added conc.hydrochloric acid (4.4 ml) at 10° C. The resulting solution was warmedto room temperature and stirred for 4.5 hours. The mixture waspulverized with ethyl acetate (800 ml) and the powder was collected byfiltration, washed with ethyl acetate and dried in vacuo. The powder(3.68 g) was suspended in a mixture of dichloromethane (12 ml) andanisole (4 ml), and trifluoroacetic acid (8 ml) was added thereto. Afterbeing stirred at room temperature for 3.5 hours, the mixture waspulverized with diisopropyl ether (800 ml) and the powder was collectedby filtration and washed with diisopropyl ether. The powder wassuspended in a mixture of water (100 ml) and ethyl acetate (30 ml) andadjusted with a saturated aqueous solution of sodium bicarbonate. Afterremoval of the insoluble materials, the aqueous layer was separated,adjusted to pH 4 with 1N hydrochloric acid and chromatographed on DiaionHP-20 (250 ml). The elution was carried out with 10% aqueous isopropylalcohol. The eluate was lyophilized to give 7β-[2-(2-aminothiazol-4-yl)-2-carboxymethoxyiminoacetamido]-3-(1-methyl-1-piperazinio)methyl-3-cephem-4-carboxylate(syn isomer) (763 mg).

IR (Nujol):1780, 1720, 1670, 1630 cm⁻¹.

NMR (D₂ O+NaHCO₃, δ):3.10 (3H, br s), 3.15-3.6 (6H, m), 4.8-5.0 (2H, m),4.53 (2H, s), 5.30 (1H, d, J=5Hz), 5.81(1H, d, J=5Hz), 6.98 (1H, s).

EXAMPLE 7

To a solution of7β-[2-(2-formamidothiazol-4-yl)-2-methoxyiminoacetamido]-3-chloromethyl-3-cephem-4-carboxylicacid (syn isomer) (920 mg) in N,N-dimethylformamide (5 ml) was added1-methyl-4-(3,4-dihydroxybenzoyl)piperazine (945 mg) at 0° C. and themixture was stirred at 0° C. for 3.5 hours. The mixture was addeddropwise to ethyl acetate (200 ml) under stirring and the pulverizedproduct was collected by filtration and washed with ethyl acetate togive7β-[2-(2-formamidothiazol-4-yl)-2-methoxyiminoacetamido]-3-[1-methyl-4-(3,4-dihydroxybenzoyl)-1-piperazinio]methyl-3-cephem-4-carboxylate(syn isomer) (1.53 g).

NMR (DMSO-d₆, δ):2.87 (3H, s), 2.9-3.05 (4H, m), 3.6-3.8 (4H, m), 3.87(3H, s), 5.16 (1H, d, J=5Hz), 5.74 (1H, dd, J=8Hz and 5Hz), 6.7-6.9 (3H,m), 7.35 (1H, s), 8.46 (1H, s), 9.1-9.5 (2H, br), 9.59 (1H, d, J=8Hz),12.50 (1H, s).

EXAMPLE 8

To a suspension of1-methyl-4-(4,5-dihydroxy-2-pyridylcarbonyl)piperazine (800 mg) indimethyl sulfoxide (8 ml) was added sodium 2-ethylhexanoate (840 mg).The solution was added dropwise a solution of7β-[2-(5-amino-1,2,4-thiadiazol-3-yl)-2-allyloxyiminoacetamido]-3-chloromethyl-3-cephem-4-carboxylatetrifluoroacetate (syn isomer) (965 mg) in dimethyl sulfoxide (7 ml) atroom temperature. After being stirred for 4 hours, the mixture waspoured into ethyl acetate (200 ml) and the resulting precipitates werecollected by filtration. The precipitates were dissolved in water andthen adjusted to pH 3.0 with aqueous sodium bicarbonate. The resultingsolution was subjected to column chromatography on Diaion HP-20 using15% aqueous isopropyl alcohol as an eluent and the objective fractionwas lyophilized to give7β-[2-(5-amino-1,2,4-thiadiazol-3-yl)-2-allyloxyiminoacetamido]-3-[1-methyl-4-(4,5-dihydroxy-2-pyridylcarbonyl)-1-piperazinio]methyl-3-cephem-4-carboxylate(syn isomer) (380 mg).

IR (Nujol):1770, 1610, 1530, 1650 cm⁻¹.

NMR (D₂ O-NaHCO₃, δ):3.16 (3H, s), 3.26-4.40 (10H), 4.78 (2H, d, J=5Hz),5.26 (1H, d, J=5Hz), 5.13-5.53 (2H, m), 5.85 (1H, dd, J=5Hz, 8Hz),5.86-6.30 (1H, m), 6.70 (1H, s), 7.70 (1H, s).

EXAMPLE 9

The following compounds were obtained according to similar manners tothose of Examples 7 and 8.

(1)7β-[2-(2-Formamidothiazol-4-yl)-2tert-butoxycarbonylmethoxyiminoacetamido]-3-[1-methyl-4-(3,4-dihydroxybenzoyl)-1-piperazinio]methyl-3-cephem-4-carboxylatehydrochloride (syn isomer)

NMR (DMSO-d₆, δ):1.40 (9H, s), 3.10 (3H, br s), 3.20-3.80 (10H), 4.55(2H, s), 4.90-5.15 (2H, m), 5.18 (1H, d, J=5Hz), 5.68 (1H, dd, J=5Hz,8Hz), 6.50-6.93 (3H, m), 6.73 (1H, s), 8.48 (1H, s),

(2) 7β-[2-(5-Amino-1,2,4-thiadiazol-3-yl)-2-ethoxyiminoacetamido]-3-[1-methyl-4-(4,5-dihydroxy-2-pyridylcarbonyl)-1-piperazinio]methyl-3-cephem-4-carboxylate(syn isomer)

IR (Nujol):3280, 1770, 1615, 1540 cm⁻¹.

NMR (DMSO-d₆, δ):1.25 (3H, t, J=7Hz), 3.06 (3H, br s), 3.3-3.8 (6H, m),4.15 (2H, q, J=7Hz), 4.2 and 5.0 (2H, m), 5.08 (1H, d, J=5Hz), 5.66 (1H,dd, J=8 and 5Hz), 7.05 (1H, s), 7.83 (1H, s), 8.04 (2H, br s), 9.41 (1H,d, J=8Hz).

EXAMPLE 10

To a suspension of7β-[2-(2-formamidothiazol-4-yl)-2-methoxyiminoacetamido]-3-[1-methyl-4-(3,4-dihydroxybenzoyl)-1-piperazinio]methyl-3-cephem-4-carboxylate(syn isomer) (1.52 g) in methanol (9 ml) was added concentratedhydrochloric acid (1.92 ml). After being stirred at room temperature for4 hours, the solution was poured into ice water (40 ml) and adjusted topH 2 with a saturated aqueous solution of sodium bicarbonate. Themixture was concentrated in vacuo to remove methanol. The concentratewas adjusted to pH 2 with a saturated aqueous solution of sodiumbicarbonate and subjected to column chromatography on "Diaion HP-20" (40ml), and the elution was carried out with 40% aqueous isopropyl alcohol.The eluate was concentrated in vacuo and lyophilized to give7β-[2-(2-aminothiazol-4-yl)-2-methoxyiminoacetamido]-3-[1-methyl-4-(3,4-dihydroxybenzoyl)-1-piperazinio]methyl-3-cephem-4-carboxylate(syn isomer) (548 mg).

IR (Nujol):1775, 1660, 1610, 1525 cm⁻¹.

NMR (DMSO-d₆, δ):3.07 (3H, s), 3.3-3.6 (4H, m), 3.7-4.0 (6H, m), 4.1 and5.05 (2H, m), 5.11 (1H, d, J=5Hz), 5.65 (1H, dd, J=8 and 5Hz), 6.71 (1H,s), 6.7-6.9 (3H, m), 9.49 (1H, d, J=8Hz).

EXAMPLE 11

The following compounds were obtained according to a similar manner tothat of Example 10.

(1)7β-[2-(2-Aminothiazol-4-yl)-2-methoxyiminoacetamido]-3-[1-methyl-4-(4,5-dihydroxy-2-pyridylcarbonyl)-1piperazinio]methyl-3-cephem-4-carboxylate(syn isomer)

IR (Nujol) 1770, 1660, 1610, 1530 cm⁻¹.

NMR (D₂ O-NaHCO₃, δ):3.20 (3H, s), 3.30-4.20 (10 H), 4.01 (3H, s), 5.36(1H, d, J=5Hz), 5.85 (1H, d, J=5Hz), 6.73 (1H, s), 6.97 (1H, s), 7.73(1H, s).

(2)7β-[2-(2-Aminothiazol-4-yl)-2-tert-butoxycarbonylmethoxyiminoacetamido]-3-[1-methyl-4-(3,4-dihydroxybenzoyl)-1-piperazinio]methyl-3-cephem-4-carboxylatedihydrochloride (syn isomer)

NMR (DMSO-d₆, δ):1.41 (9H, s), 3.15 (3H, s), 3.20-4.30 (10H), 4.63 (2H,s), 5.33 (1H, d, J=5Hz), 5.88 (1H, dd, J=8Hz, 5Hz), 6.63-6.95 (3H, m),6.95 (1H, s), 9.80 (1H, d, J=8Hz).

EXAMPLE 12

To a solution of 4,5-dihydroxy-2-pyridinecarboxylic acid (3.1 g) and1-hydroxybenzotriazole (4.0 g) in dimethyl sulfoxide (30 ml) was addedN,N'-dicyclohexylcarbodiimide (6.2 g). After being stirred at roomtemperature for 1 hour, the reaction mixture was added to a solution of7β-[2-(5-amino-1,2,4-thiadiazol-3-yl)-2-methoxyiminoacetamido]-3-(1-methyl-1-piperazinio)methyl-3-cephem-4-carboxylate(syn isomer) (10.0 g) and N-trimethylsilylacetamide (3.0 g) in dimethylsulfoxide (50 ml), and the stirring was continued for 30 minutes. Themixture was added dropwise to diisopropyl ether (1.5 l) and theresulting precipitate was collected by filtration, which was dissolvedin an aqueous solution of sodium bicarbonate. After the insolublematerial was filtered off, the filtrate was adjusted to pH 3.0 with 1Nhydrochloric acid and subjected to column chromatography on "DiaionHP-20" with 5% aqueous isopropyl alcohol as an eluent. The eluate waslyophilized to give7β-[2-(5-amino-1,2,4-thiadiazol-3-yl)-2-methoxyiminoacetamido]-3-[1-methyl-4-(4,5-dihydroxy-2-pyridylcarbonyl)-1-piperazinio]methyl-3-cephem-4-carboxylate(syn isomer) (0.5 g).

IR (Nujol):3300, 1770, 1610 cm⁻¹.

NMR (DMSO-d₆, δ):3.08 (3H, s), 3.92 (3H, s), 5.12 (1H, d, J=5Hz), 5.70(1H, dd, J=5, 8Hz), 7.10 (1H, s , 7.89 (1H, s), 8.10 (2H, br s), 9.47(1H, d, J=8Hz).

EXAMPLE 13

The solution of7B-[2-(5-amino-1,2,4-thiadiazol-3-yl)-2-carboxymethoxyiminoacetamido]-3-(1-methyl-1-piperazinio)methyl-3-cephem-4-carboxylate(syn isomer) (540 mg) in acetone (5 ml) and water (5 ml) was adjusted topH 7.0 with a saturated aqueous solution of sodium bicarbonate. To themixture was added 3,4-diacetoxybenzoyl chloride (308 mg) at pH 6.5-7.0.The mixture was stirred for an hour and evaporated in vacuo to removeacetone. The residue was adjusted to pH 8.5 with a saturated aqueoussolution of sodium bicarbonate and then stirred for 2 hours at roomtemperature. The mixture was adjusted to pH 3.5 with 3N hydrochloricacid and then filtered. The filtrate was subjected to columnchromatography on Diaion HP-20 using 5% aqueous isopropyl alcohol as aneluent. The eluate was evaporated in vacuo to remove isopropyl alcoholand then lyophilized to give7β-[2-(5-amino-1,2,4-thiadiazol-3-yl)-2carboxymethoxyiminoacetamido]-3-[1-methyl-4-(3,4-dihydroxybenzoyl)- 1-piperazinio]methyl-3-cephem-4-carboxylate(syn isomer) (265 mg).

IR (Nujol):3300, 1770, 1600, 1620 cm⁻¹.

NMR (D₂ O, δ):3.15 (3H, br s), 5.32 (1H, d, J=5Hz), 5.85 (1H, d, J=5Hz),6.90 (3H, br s).

EXAMPLE 14

To a solution of7β-[2-(5-amino-1,2,4-thiadiazol-3-yl)-2-ethoxyiminoacetamido]-3-(1-methyl-1-piperazinio)methyl-3-cephem-4-carboxylate(syn isomer) (255 mg) in a mixture of water (5 ml) and acetone (5 ml)was added 4,5-diacetoxy-2-pyridinecarbonyl chloride hydrochloride (309mg), with maintaining the pH of the solution to 8.0-8.5 by addition ofsaturated aqueous sodium bicarbonate. After stirring for 30 minutes,acetone was removed by evaporation in vacuo and the residue was adjustedto pH 8 and stirred for 2 hours. The mixture was adjusted to pH 3.0 with1N hydrochloric acid and chromatographed on Diaion HP-20 (50 ml), andthe elution was carried out with 10% aqueous isopropyl alcohol. Theeluate was lyophilized to give7β-[2-(5-amino-1,2,4-thiadiazol-3-yl)-2-ethoxyiminoacetamido]-3-[1-methyl-4-(4,5-dihydroxy-2-pyridylcarbonyl)-1-piperazinio]methyl-3-cephem-4-carboxylate(syn isomer) (155 mg).

IR (Nujol):3200, 1765, 1610, 1520 cm⁻¹.

NMR (DMSO-d₆, δ):1.24 3H, t, J=7Hz), 3.05 (3H, br s), 3.3-3.9 (10H, m),4.13 (2H, q, J=7Hz), 4.1 and 4.96 (2H, m), 5.09 (1H, d, J=5Hz), 5.67(1H, dd, J=8 and 5Hz), 7.04 (1H, s), 7.84 (1H, s), 8.03 (2H, br s), 9.43(1H, d, J=8Hz).

EXAMPLE 15

The following compounds were obtained according to similar manners tothose of Examples 13 and 14.

(1)7β-[2-(2-Formamidothiazol-4-yl)-2-methoxyiminoacetamido]-3-[1-methyl-4-(4,5-dihydroxy-2-pyridylcarbonyl)-1-piperazinio]methyl-3-cephem-4-carboxylate(syn isomer)

NMR (D₂ O-NaHCO₃, δ):3.22 (3H, s), 3.30-4.30 (10H), 4.05 (3H, s), 5.38(1H, d, J=5Hz), 5.86 (1H, d, J=5Hz), 6.73 (1H, s), 7.46 (1H, s), 7.73(1H, s), 8.53 (1H, s) ,

(2)7β-[2-(5-Amino-1,2,4-thiadiazol-3-yl)-2carboxymethoxyiminoacetamido]-3-[1-methyl-4-(4,5-dihydroxy-2-pyridylcarbonyl)-1-piperazinio]methyl-3-cephem-4-carboxylate(syn isomer)

IR (Nujol):3300, 1780, 1660, 1600 cm⁻¹.

NMR (D₂ O+NaHCO₃, δ):3.33 (3H, s), 5.35 (1H, d, J=5Hz), 5.88 (1H, d,J=5Hz), 6.73 (1H, s), 7.73 (1H, s).

(3)7β-[2-(2-Aminothiazol-4-yl)-2-carboxymethoxyiminoacetamido]-3-[1-methyl-4-(4,5-dihydroxy-2-pyridylcarbonyl)-1-piperazinio]methyl-3-cephem-4-carboxylate(syn

IR (Nujol):1770, 1660, 1610, 1525 cm⁻¹.

NMR (D₂ O-NaHCO₃, δ):3.16 (3H, s), 3.52 (2H, br s), 3.20-4.30 (8H),4.50-5.00 (2H +2H, m), 5.32 (1H, d, J=5Hz), 5.85 (1H, d, J=5Hz), 6.70(1H, s), 6.98 (1H, s), 7.70 (1H, s),

EXAMPLE 16

To a suspension of7β-[2-(2-aminothiazol-4-yl)-2-tert-butoxycarbonylmethoxyiminoacetamido]-3-[1-methyl-4-(3,4-dihydroxybenzoyl)-1-piperazinio]methyl-3-cephem-4-carboxylatedihydrochloride (syn isomer) (1.35 g) in dichloromethane (5.4 ml) andanisole (1.3 ml) was added dropwise trifluoroacetic acid (4 ml) at 25°C. The stirring was continued for 3 hours at the same temperature. Thereaction mixture was poured into diisopropyl ether and the resultingprecipitates were collected by filtration. The precipitates weredissolved in water and then adjusted to pH 3.0 with aqueous sodiumbicarbonate. The resulting solution was subjected to columnchromatography on Diaion HP-20 using 15% aqueous isopropyl alcohol as aneluent. The object fractions were lyophilized to give7δ-[2-(2-aminothiazol-4-yl)-2-carboxymethoxyiminoacetamido]-3-[1-methyl-4-(3,4-dihydroxybenzoyl)-1-piperazinio]methyl-3-cephem-4-carboxylate(syn isomer) (320 mg).

IR (Nujol):1770, 1660, 1610, 1520 cm⁻¹.

NMR (D₂ O-NaHCO₃, δ):3.15 (3H, s), 3.20-4.30 (10H), 4.55 (2H, s),4.60-5.00 (2H, m), 5.32 (1H, d, J=5Hz), 5.82 (1H, d, J=5Hz), 6.80-7.10(3H, m), 6.90 (1H, s).

EXAMPLE 17

To 1N hydrochloric acid was added7β-[2-(5-amino-1,2,4-thiadiazol-3-yl)-2-(1-carboxy-1-methylethoxyimino)acetamido]-3-[N,N-dimethyl-N-((5-hydroxy-4-oxo-1,4-dihydropyridin-2-yl)methyl}ammonio]methyl-3-cephem-4-carboxylate(syn isomer) (2.2 g). The mixture was stirred under ice-cooling for 3hours. The resulting precipitate was collected by filtration, washedwith a small amount of cold water and dried under reduced pressure togive7β-[2-(5-amino-1,2,4-thiadiazol-3-yl)-2-(1-carboxy-1-methylethoxyimino)acetamido]-3-{N,N-dimethyl-N-{(5-hydroxy-4-oxo-1,4-dihydropyridin-2-yl)methyl}ammonio]methyl-3-cephem-4-carboxylatehydrochloride (syn isomer) (1.1 g).

IR (Nujol):3450-3150 (br), 2650, 1770, 1670, 1610 cm⁻¹.

EXAMPLE 18

To a mixture of water (22 ml) and acetone (22 ml) was added7β-[2-(5-amino-1,2,4-thiadiazol-3-yl)-2-(1-carboxy-1-methylethoxyimino)acetamido]-3-[N,N-dimethyl-N-{(5-hydroxy-4-oxo-1,4-dihydropyridin-2-yl)methyl}ammonio]methyl-3-cephem-4-carboxylate(syn isomer) (2.20 g). To the resulting solution was added 97% sulfuricacid (0.699 g). The solution was stirred for 6.5 hours at 25° to 30° C.The resulting crystals were collected by filtration, washed with amixture of water (9 ml) and acetone (9 ml) and dried under reducedpressure to give sulfuric acid salt (2.17 g) of7β-[2-(5-amino-1,2,4-thiadiazol-3-yl)-2-(1-carboxy-1-methylethoxyimino)acetamido]-3-[N,N-dimethyl-N-{(5-hydroxy-4-oxo-1,4,dihydropyridin-2-yl)methyl}ammonio]-methyl-3-cephem-4-carboxylate(syn isomer).

mp:145 (dec.).

IR (Nujol):3450-3150 (br), 2650 (br), 1780, 1692, 1615, 1558, 1529cm⁻¹.

EXAMPLE 19

The following compounds were obtained according to similar manner tothose of Examples 5 and 6.

(1)7β-[2-(5-Amino-1,2,4-thiadiazol-3-yl)-2-methoxyiminoacetamido]-3-[N,N-dimethyl-N-{(5-hydroxy-4-oxo-1,4-dihydropyridin-2-yl)methyl}ammonio]methyl-3-cephem-4-carboxylate(syn isomer)

IR (Nujol):3250 (br), 1765, 1650, 1600 cm⁻¹.

(2)7β-[2-[2-Formamidothiazol-4-yl)-2-tert-butoxycarbonylmethoxyiminoacetamido]-3-[N,N-dimethyl-N-{(5-hydroxy-4-oxo-1,4-dihydropyridin-2-yl)methyl}ammonio]-methyl-3-cephem-4-carboxylate(syn isomer)

IR (Nujol):3300 (br), 1780, 1730, 1675, 1613 cm⁻¹.

(3)7β-[2-(2-Formamidothiazol-4-yl)-2-methoxyiminoacetamido]-3-[N,N-dimethyl-N-{2-(5-hydroxy-4-oxo-1,4-dihydropyridin-2-yl)ethyl}ammonio]methyl-3-cephem-4-carboxylate(syn isomer)

IR (Nujol):3200 (br) 1772, 1670, 1608 cm⁻¹.

(4) 7β-[2-(5-Amino-1,2,4-thiadiazol-3-yl-2-methoxyiminoacetamido]-3-[1-{2-(5-hydroxy-4-oxo-1,4-dihydropyridin-2-yl)ethyl)-1-pyrrolidinio]methyl-3-cephem-4-carboxylate(syn isomer)

IR (Nujol):3300 (br), 1765, 1663 (sh), 1620 cm⁻¹.

(5)7β-[2-(5-Amino-1,2,4-thiadiazol-3-yl)-2-(1-carboxy-1-methylethoxyimino)acetamido]-3-[1-{2-(5-hydroxy-4-oxo-1,4-dihydropyridin-2-yl)ethyl}-1-pyrrolidinio]methyl-3-cephem-4-carboxylate(syn isomer)

IR (Nujol):3300 (br), 1770, 1670˜1620 (br) cm⁻¹.

(6)7β-[2-(5-Amino-1,2,4-thiadiazol-3-yl)-2-(1-carboxy-1-methylethoxyimino)acetamido]-3-[N,N-dimethyl-N-{2-(5-hydroxy-4-oxo-1,4-dihydropyridin-2-yl)ethyl}ammonio]-methyl-3-cephem-4-carboxylate(syn isomer)

IR (Nujol):3300 (br), 1775, 1670, 1615 cm⁻¹.

(7)7β-[2-(5-Amino-1,2,4-thiadiazol-3-yl)-2-methoxyiminoacetamido]-3-[N,N-dimethyl-N-{2-(5-hydroxy-4-oxo-1,4-dihydropyridin-2-yl)ethyl}ammonio]methyl-3-cephem-4-carboxylate(syn isomer)

IR (Nujol):3250 (br), 1770, 1670 (sh), 1610 cm⁻¹.

(8)7β-[2-(2-Formamidothiazol-4-yl)-2-(1-tert-butoxycarbonyl-1-methylethoxyimino)acetamido]-3-[N,N-dimethyl-N-{2-(5-hydroxy-4-oxo-1,4-dihydropyridin-2-yl)ethyl}ammonio]methyl-3-cephem-4-carboxylate(syn isomer)

IR (Nujol):3200, 1770, 1720, 1685 (sh), 1668, 1605 cm⁻¹.

(9)7β-[2-(2-Formamidothiazol-4-yl)-2-(1-tert-butoxycarbonyl-1-methylethoxyimino)acetamido]-3-[N,N-dimethyl-N-{(5-hydroxy-4-oxo-1,4-dihydropyridin-2-yl)methyl}ammonio]methyl-3-cephem-4-carboxylate(syn isomer)

IR (Nujol):3250, 1780, 1720, 1675, 1611 cm⁻¹.

(10)7β-[2-(2-Aminothiazol-4-yl)-2-tert-butoxycarbonylmethoxyiminoacetamido]-3-[N,N-dimethyl-N-{(5-hydroxy-4-oxo-1,4-dihydropyridin-2-yl)methyl}ammonio]methyl-3-cephem-4-carboxylate(syn isomer)

IR (Nujol):3300 br), 1750, 1730 (sh), 1667, 1608 cm⁻¹.

(11)7β-[2-(2-Aminothiazol-4-yl)-2-methoxyiminoacetamido]-3-{N,N-dimethyl-N-{2-(5-hydroxy-4-oxo-1,4-dihydropyridin-2-yl)ethyl}ammonio]methyl-3-cephem-4-carboxylate(syn isomer)

IR (Nujol):3250 (br), 1770, 1660, 1705 cm⁻¹.

(12)7β-[2-(2-Aminothiazol-4-yl)-2-(1-tert-butoxycarbonyl-1-methylethoxyimino)acetamido]-3-[N,N-dimethyl-N-{2-(5-hydroxy-4-oxo-1,4-dihydropyridin-2-yl)ethyl}-ammonio]methyl-3-cephem-4-carboxylate(syn isomer)

IR (Nujol):3250 br , 1770, 1720, 1662, 1608 cm⁻¹.

(13)7β-[2-(2-Aminothiazol-4-yl)-2-(1-tert-butoxycarbonyl-1-methylethoxyimino)acetamido]-3-[N,N-dimethyl-N{(5-hydroxy-4-oxo-1,4-dihydropyridin-2-yl)methyl}ammonio]methyl-3-cephem-4-carboxylate(syn isomer)

IR (Nujol):3350 (br), 1775, 1720, 1672, 1608 cm⁻¹.

(14)7β-[2-(2-Aminothiazol-4-yl}-2-carboxymethoxyiminoacetamido]-3-[N,N-dimethyl-N-{(5-hydroxy-4-oxo-1,4-dihydropyridin-2-yl)methyl}ammonio]methyl-3-cephem-4-carboxylate(syn isomer)

IR (Nujol 3250 br), 1770, 1662 (sh), 1600 cm⁻¹.

(15)7β-[2-(2-Aminothiazol-4-yl)-2-(1-carboxy-1-methylethoxyimino)acetamido]-3-[N,N-dimethyl-N-{2-(5-hydroxy-4-oxo-1,4-dihydropyridin-2-yl)ethyl}ammonio]methyl-3-cephem-4-carboxylate(syn isomer)

IR (Nujol):3250 (br), 1770, 1665, 1610 cm⁻¹.

(16)7β-[2-(2-Aminothiazol-4-yl)-2-(1-carboxy-1-methylethoxyimino)acetamido]-3-[N,N-dimethyl-N-{(5-hydroxy-4-oxo-1,4-dihydropyridin-2-yl)methyl}ammonio]-methyl-3-cephem-4-carboxylate(syn isomer)

IR (Nujol):3250, 1770, 1662, 1607 cm⁻¹.

(17)7β-[2-(5-Amino-1,2,4-thiadiazol-3-yl)-2-(1-carboxy-1-methylethoxyimino)acetamido]-3-[N,N-dimethyl-N-{(5-hydroxy-4-oxo-1,4-dihydropyridin-2-yl)methyl}ammonio]-methyl-3-cephem-4-carboxylate hydrochloride (syn isomer)

IR (Nujol):3450-3150 (br), 2650, 1770, 1670, 1610 cm⁻¹.

(18) Sulfuric acid salt of7β-[2-(5-amino-1,2,4-thiadiazol-3-yl)-2-(1-carboxy-1-methylethoxyimino)acetamido]-3-[N,N-dimethyl-N-{(5-hydroxy-4-oxo-1,4-dihydropyridin-2-yl)methyl}ammonio]methyl-3-cephem-4-carboxylate(syn isomer)

IR (Nujol):3450-3150 (br), 2650 (br), 1780, 1692, 1615, 1558, 1529 cm⁻¹.

(19)7β-[2-(5-Amino-1,2,4-thiadiazol-3-yl)-2-(1-carboxy-1-methylethoxyimino)acetamido]-3-[1-methyl-4-(4,5-dihydroxy-2-pyridylcarbonyl)-1-piperazinio]methyl-3-cephem-4-carboxylate(syn isomer)

IR (Nujol):1770 cm⁻¹.

(20)7β-[2-(2-Formamidothiazol-4-yl)-2-methoxyiminoacetamido]-3-[1-methyl-4-(3,4-dihydroxybenzoyl)-1-piperazinio]methyl-3-cephem-4-carboxylate(syn isomer)

NMR (DMSO-d₆, δ):2.87 (3H, s), 2.9-3.05 (4H, m), 3.6-3.8 (4H, m), 3.87(3H, s), 5.16 (1H, d, J=5Hz) 5.74 (1H, dd, J=8Hz and 5Hz), 6.7-6.9 (3H,m), 7.35 (1H, s), 8.46 (1H, s), 9.1-9.5 (2H, br), 9.59 (1H, d, J=8Hz),12.50 (1H, s).

(21)7β-[2-(5-Amino-1,2,4-thiadiazol-3-yl)-2-allyloxyiminoacetamido]-3-[1-methyl-4-(4,5-dihydroxy-2-pyridylcarbonyl)-1-piperazinio]methyl-3-cephem-4-carboxylate(syn isomer)

IR (Nujol):1770, 1610, 1530, 1650 cm⁻¹.

(22)7β-[2-(2-Formamidothiazol-4-yl)-2-tert-butoxycarbonylmethoxyiminoacetamido]-3-[1-methyl-4-(3,4-dihydroxybenzoyl)-1-piperazinio]methyl-3-cephem-4-carboxylate hydrochloride (syn isomer)

NMR DMSO-d₆, δ):1.40 (9H, s), 3.10 (3H, br s), 3.20-3.80 (10H), 4.55(2H, s), 4.90-5.15 (2H, m), 5.18 (1H, d, J=5Hz), 5.68 (1H, dd, J=5Hz,8Hz), 6.50-6.93 (3H, m), 6.73 (1H, s), 8.48 (1H, s) .

(23)7β-[2-(5-Amino-1,2,4-thiadiazol-3-yl)-2-ethoxyiminoacetamido]-3-[1-methyl-4-(4,5-dihydroxy-2-pyridylcarbonyl)-1-piperazinio]methyl-3-cephem-4-carboxylate(syn isomer)

IR (Nujol):3280, 1770, 1615, 1540 cm⁻¹.

(24)7β-[2-(2-Aminothiazol-4-yl)-2-methoxyiminoacetamido]-3-[1-methyl-4-(3,4-dihydroxybenzoyl)-1-piperazinio]methyl-3-cephem-4-carboxylate(syn isomer)

IR (Nujol):1775, 1660, 1610, 1525 cm⁻¹.

(25)7β-[2-(2-Aminothiazol-4-yl)-2-methoxyiminoacetamido]-3-[1-methyl-4-(4,5-dihydroxy-2-pyridylcarbonyl)-1-piperazinio]methyl-3-cephem-4-carboxylate(syn isomer)

IR (Nujol):1770, 1660, 1610, 1530 cm⁻¹.

(26)7β-[2-(2-Aminothiazol-4-yl)-2-tert-butoxycarbonylmethoxyiminoacetamido]-3-[1-methyl-4-(3,4-dihydroxybenzoyl)-1-piperazinio]methyl-3-cephem-4-carboxylatedihydrochloride (syn isomer)

NMR (DMSO-d₆, δ):1.41 (9H, s), 3.15 (3H, s), 3.20-4.30 (10H), 4.63 2H,s), 5.33 (1H, d, J=5Hz), 5.88 (1H, dd, J=8Hz, 5Hz), 6.63-6.95 (3H, m),6.95 (1H, s), 9.80 (1H, d, J=8Hz).

(27)7β-[2-(5-Amino-1,2,4-thiadiazol-3-yl)-2-methoxyiminoacetamido]-3-[1-methyl-4-(4,5-dihydroxy-2-pyridylcarbonyl)-1-piperazinio]methyl-3-cephem-4-carboxylate (syn isomer)

IR (Nujol):3300, 1770, 1610 cm⁻¹.

(28)7β-[2-(5-Amino-1,2,4-thiadiazol-3-yl)-2-carboxymethoxyiminoacetamido]-3-[1-methyl-4-(3,4-dihydroxybenzoyl)-1-piperazinio]methyl-3-cephem-4-carboxylate(syn isomer)

IR (Nujol):3300, 1770, 1600, 1620 cm⁻¹.

(29)7β-[2-(2-Formamidothiazol-4-yl)-2-methoxyiminoacetamido]-3-[1-methyl-4-(4,5-dihydroxy-2-pyridylcarbonyl)-1-piperazinio]methyl-3-cephem-4-carboxylate(syn isomer)

NMR (D₂ O-NaHCO₃, δ):3.22 (3H, s), 3.30-4.30 (10H), 4.05 (3H, s), 5.38(1H, d, J=5Hz), 5.86 (1H, d, J=5Hz), 6.73 (1H, s), 7.46 (1H, s), 7.73(1H, s), 8.53 (1H, s).

(30)7β-[2-(5-Amino-1,2,4-thiadiazol-3-yl)-2-carboxymethoxyiminoacetamido]-3-[1-methyl-4-(4,5-dihydroxy-2-pyridylcarbonyl)-1-piperazinio]methyl-3-cephem-4-carboxylate(syn isomer)

IR (Nujol):3300, 1780, 1660, 1600 cm⁻¹.

(31)7β-[2-(2-Aminothiazol-4-yl)-2-carboxymethoxyiminoacetamido]-3-[1-methyl-4-(4,5-dihydroxy-2-pyridylcarbonyl)-1-piperazinio]methyl-3-cephem-4-carboxylate(syn isomer)

IR (Nujol):1770, 1660, 1610, 1525 cm⁻¹.

(32)7β-[2-(2-Aminothiazol-4-yl)-2-carboxymethoxyiminoacetamido]-3-[1-methyl-4-(3,4-dihydroxybenzoyl)-1-piperazinio]methyl-3-cephem-4-carboxylate(syn isomer)

IR (Nujol):1770, 1660, 1610, 1520 cm⁻¹.

EXAMPLE 20

The following compounds were obtained according to similar manners tothose of Examples 7 and 8.

(1)7β-[2-(5-Amino-1,2,4-thiadiazol-3-yl)-2-(1-carboxy-1-methylethoxyimino)acetamido]-3-[1-methyl-4-(4,5-dihydroxy-2-pyridylcarbonyl)-1-piperazinio]methyl-3-cephem-4-carboxylate(syn isomer)

IR (Nujol):1770 cm⁻¹.

NMR (D₂ O+NaHCO₃, δ):1.55 (6H, s , 3.16 (3H, s), 3.2-5.1 (12H, m), 5.35(1H, d, J=5Hz), 5.85 (1H, d, J=5Hz), 6.70 (1H, s), 7.70 (1H, s).

(2)7β-[2-(2-Aminothiazol-4-yl)-2-methoxyiminoacetamido]-3-[1-methyl-4-(3,4-dihydroxybenzoyl)-1-piperazinio]methyl-3-cephem-4-carboxylate(syn isomer)

IR (Nujol):1775, 1660, 1610, 1525 cm⁻¹.

(3)7β-[2-(2-Aminothiazol-4-yl)-2-methoxyiminoacetamido]-3-[1-methyl-4-(4,5-dihydroxy-2-pyridylcarbonyl)-1piperazinio]methyl-3-cephem-4-carboxylate(syn isomer)

IR (Nujol):1770, 1660, 1610, 1530 cm⁻¹.

(4)7β-[2-(2-Aminothiazol-4-yl)-2-tert-butoxycarbonylmethoxyiminoacetamido]-3-[1-methyl-4-(3,4-dihydroxybenzoyl)-1-piperazinio]methyl-3-cephem-4-carboxylatedihydrochloride (syn isomer)

NMR [DMSO-d₆, δ):1.41 (9H, s), 3.15 (3H, s), 3.20-4.30 (10H),4.63 (2H,s), 5.33 (1H, d, J=5Hz), 5.88 (1H, dd, J=8Hz, 5Hz), 6.63-6.95 (3H, m),6.95 (1H, s), 9.80 (1H, d, J=8Hz) ,

(5)7β-[2-(5-Amino-1,2,4-thiadiazol-3-yl)-2-methoxyiminoacetamido]-3-[1-methyl-4-(4,5-dihydroxy-2-pyridylcarbonyl)-1-piperazinio]methyl-3-cephem-4-carboxylate(syn isomer)

IR (Nujol):3300, 1770, 1610 cm⁻¹.

(6)7β-[2-(5-Amino-1,2,4-thiadiazol-3-yl)-2-carboxymethoxyiminoacetamido]-3-[1-methyl-4-(3,4-dihydroxybenzoyl)-1-piperazinio]methyl-3-cephem-4-carboxylate(syn isomer)

IR (Nujol):3300, 1770, 1600, 1620 cm⁻¹.

(7)7β-[2-(2-Formamidothiazol-4-yl)-2-methoxyiminoacetamido]-3-[1-methyl-4-(4,5-dihydroxy-2-pyridylcarbonyl)-1-piperazinio]methyl-3-cephem-4-carboxylate(syn isomer)

NMR (D₂ O-NaHCO₃, δ):3.22 (3H, s), 3.30-4.30 (10H), 4.05 (3H, s), 5.38(1H, d, J=5Hz), 5.86 (1H, d, J=5Hz), 6.73 (1H, s), 7.46 (1H, s), 7.73(1H, s), 8.53 (1H, s) .

(8)7β-[2-(5-Amino-1,2,4-thiadiazol-3-yl)-2-carboxymethoxyiminoacetamido]-3-[1-methyl-4-(4,5-dihydroxy-2-pyridylcarbonyl)-1-piperazinio]methyl-3-cephem-4-carboxylate(syn isomer)

IR (Nujol):3300, 1780, 1660, 1600 cm⁻¹.

(9)7β-[2-(2-Aminothiazol-4-yl)-2-carboxymethoxyiminoacetamido]-3-[1-methyl-4-(4,5-dihydroxy-2-pyridylcarbonyl)-1-piperazinio]methyl-3-cephem-4-carboxylate(syn isomer)

IR (Nujol):1770, 1660, 1610, 1525 cm⁻¹.

(10)7β-[2-(2-Aminothiazol-4-yl)-2-carboxymethoxyiminoacetamido]-3-[1-methyl-4-(3,4-dihydroxybenzoyl)-1-piperazinio]methyl-3-cephem-4-carboxylate[syn isomer)

IR (Nujol):1770, 1660, 1610, 1520 cm⁻¹.

What we claimed is:
 1. A compound of the formula: ##STR17## wherein R¹is amino or a protected amino group,Z is N or CH, R² is lower alkyl,mono(or di or tri)halo(lower)alkyl, lower alkenyl, lower alkynyl,phenyl, naphthyl, phenyl(lower)alkyl, carboxy(lower)alkyl or protectedcarboxy(lower)alkyl, and R is a group of the formula: ##STR18## in whichR⁶ is lower alkyl, R⁷ and R⁸ are each hydroxy or a protected hydroxygroup, and Y is N or CH, or a pharmaceutically acceptable salt thereof.2. A compound of claim 1,wherein R¹ is amino or acylamino and R² islower alkyl, lower alkenyl, carboxy(lower)alkyl or protectedcarboxy(lower)alkyl.
 3. A compound of claim 2,wherein R¹ is amino orlower alkanoylamino and R² is lower alkyl, lower alkenyl,carboxy(lower)alkyl or esterified carboxy(lower)alkyl.
 4. A compound ofclaim 1,wherein R¹ is amino, Z is N and R² is carboxy(lower)alkyl.
 5. Anantimicrobial pharmaceutical composition which comprises, as an activeingredient, an effective amount of a compound of claim 1 or apharmaceutically acceptable salt thereof in admixture with apharmaceutically acceptable carrier.
 6. A method for the treatment ofinfectious diseases which comprises administering an effective amount ofa compound of claim 1 or a pharmaceutically acceptable salt thereof to ahuman or animal.
 7. A compound of the formula: ##STR19## wherein R¹ isamino or a protected amino group,Z is N, R² is lower alkyl, mono(or dior tri)halo(lower)alkyl, lower alkenyl, lower alkynyl, phenyl, naphthyl,phenyl(lower)alkyl, carboxy(lower)alkyl or protectedcarboxy(lower)alkyl, and R is a group of the formula: ##STR20## in whichR⁶ is lower alkyl, R⁷ and R⁸ are each hydroxy or a protected hydroxygroup, and Y is N or CH, or a pharmaceutically acceptable salt thereof.8. A compound of the formula: ##STR21## wherein R¹ is amino or aprotected amino group,Z is CH, R² is lower alkyl, mono(or di ortri)halo(lower)alkyl, lower alkenyl, lower alkenyl, phenyl, naphthyl,phenyl(lower)alkyl, carobxy(lower)alkyl or protectedcarboxy(lower)alkyl, and R is or a group of the formula: ##STR22## inwhich R⁶ is lower alkyl, R⁷ and R⁸ are each hydroxy or a protectedhydroxy group, and Y is N or CH, or a pharmaceutically acceptable saltthereof.
 9. The compound of claim 8, wherein is a group of the formula:##STR23##
 10. The compound of claim 9, wherein Y is CH.
 11. The compoundof claim 9, wherein Y is N.
 12. The compound of claim 8, wherein R¹ isamino and each of R⁵, R⁷ and R⁸ are hydroxy.